Gravity spun polycaprolactone fibres for soft tissue engineering:interaction with fibroblasts and myoblasts in cell culture

Poly(ε-caprolactone) (PCL) fibres were produced by wet spinning from solutions in acetone under low shear (gravity flow) conditions. As-spun PCL fibres exhibited a mean strength and stiffness of 7.9 MPa and 0.1 GPa, respectively and a rough, porous surface morphology. Cold drawing to an extension of 500% resulted in increases in fibre strength (43 MPa) and stiffness (0.3 GPa) and development of an oriented, fibrillar surface texture. The proliferation rate of Swiss 3T3 mouse fibroblasts and C2C12 mouse myoblasts on as-spun, 500% cold-drawn and gelatin-modified PCL fibres was determined in cell culture to provide a basic measure of the biocompatibility of the fibres. Proliferation of both cell types was consistently higher on gelatin-coated fibres relative to as-spun fibres at time points below 7 days. Fibroblast growth rates on cold-drawn PCL fibres exceeded those on as-spun fibres but myoblast proliferation was similar on both substrates. After 1 day in culture, both cell types had spread and coalesced on the fibres to form a cell layer, which conformed closely to the underlying topography. The high fibre compliance combined with a potential for modifying the fibre surface chemistry with cell adhesion molecules and the surface architecture by cold drawing to enhance proliferation of fibroblasts and myoblasts, recommends further investigation of gravity-spun PCL fibres for 3-D scaffold production in soft tissue engineering. © 2005 Elsevier Ltd. All rights reserved.

Simplified recording of soft contact lens fit

Purpose: To determine the critical fitting characteristics of modern soft contact lens fits and from this to devise a simplified recording scheme. Methods: Ten subjects (aged 28.1 ± 7.4 years) wore eight different modern soft contact lenses. Video was captured and analysed of blink (central and up-gaze), excursion lag (up, down, right and left gaze) and push-up movement, centration and coverage. Results: Lens centration was on average close to the corneal centre. Movement on blink was significantly smaller in up-gaze than in primary-gaze (p<0.001). Lag was greatest in down-gaze and least in up-gaze (p<0.001). Push-up test recovery speed was 1.32±0.73mm/s. Overall lens movement was determined best by assessing horizontal lag, movement on blink in up-gaze and push-up recovery speed. Steeper lens base-curves did not have a significant effect on lens fit characteristics. Contact lens material did influence lens fit characteristics, particularly silicone-hydrogels which generally had lower centration and a faster push-up speed of recovery than HEMA lenses (p<0.05). Conclusion: Lag on vertical gaze, and movement on blink in primary gaze generally provide little extra information on overall lens movement compared to horizontal lag, movement on blink in up gaze and push-up recovery speed. They can therefore be excluded from a simplified recording scheme. A simplified and comprehensive soft contact lens fit recording system could consist of a cross-hairs indicating the centre of the cornea; a circle to indicate the lens centration; a mark on the relevant position of the circle to indicate any limbal incursion; a grade (‘B’) below for movement with blink in up-gaze, a grade (‘L’) to the side for horizontal lag and a grade above (‘P’) for the assessed push-up recovery speed.

The influence of corneoscleral topography on soft contact lens fit

To evaluate the influence of peripheral ocular topography, as evaluated by optical coherence tomography (OCT), compared with traditional measures of corneal profile using keratometry and videokeratoscopy, on soft contact lens fit.

A generative probabilistic oriented wavelet model for texture segmentation

This Letter addresses image segmentation via a generative model approach. A Bayesian network (BNT) in the space of dyadic wavelet transform coefficients is introduced to model texture images. The model is similar to a Hidden Markov model (HMM), but with non-stationary transitive conditional probability distributions. It is composed of discrete hidden variables and observable Gaussian outputs for wavelet coefficients. In particular, the Gabor wavelet transform is considered. The introduced model is compared with the simplest joint Gaussian probabilistic model for Gabor wavelet coefficients for several textures from the Brodatz album [1]. The comparison is based on cross-validation and includes probabilistic model ensembles instead of single models. In addition, the robustness of the models to cope with additive Gaussian noise is investigated. We further study the feasibility of the introduced generative model for image segmentation in the novelty detection framework [2]. Two examples are considered: (i) sea surface pollution detection from intensity images and (ii) image segmentation of the still images with varying illumination across the scene.

Objective clinical performance of ‘comfort-enhanced' daily disposable soft contact lenses

Purpose: To examine the objective clinical performance of ‘comfort-enhanced’ daily disposable contact lenses over a 16-h day. Methods: Four contact lenses (Hilafilcon B, Etafilcon A Plus, Nelfilcon A and Nelfilcon A Plus) were evaluated in an investigator masked, open label trial at the end of a week’s bilateral wear. Pre-lens noninvasive tear break-up time (PL-NITBUT), tear prism height, bulbar hyperaemia and ocular surface temperature (OST) were measured with the lens in situ at 8, 12 and 16 h of wear. Results: There was no difference between how many hours the lenses types were worn each day (F = 0.90, p = 0.44). The PL-NITBUT decreased with the duration of daily lens wear (F = 32.0, p < 0.001) and was more stable with Nelfilcon A Plus (F = 6.00, p = 0.002) than with the other lenses evaluated. Bulbar blood vessels increased in coverage (F = 11.5, p < 0.001) but not overall redness (F = 0.0, p = 0.99) with the duration of daily lens wear, but there was no difference between the lenses (p > 0.05). The tear prism height decreased with the duration of daily wear (F = 27.0, p < 0.001) and differed between lenses (F = 2.9, p = 0.04). The OST decreased with the duration of lens wear (F = 119.7, p < 0.001) and was reduced by daily disposable lens wear (F = 7.88, p < 0.001), but did not differ between lenses (F = 0.88, p = 0.45). Conclusions: Objective measures of tear film indicated a difference between the lenses evaluated for PLNITBUT and tear prism height, but not for wearing time or bulbar conjunctival hyperaemia. Therefore clinical benefits of daily disposable ‘comfort enhancing’ contact lenses can be measured, but challenges remain in producing contact lenses that do not compromise anterior eye physiology over the whole day. 2010 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

The role of texture amplitude in shape from shading: evidence from a haptic matching task

The pattern of illumination on an undulating surface can be used to infer its 3-D form (shape from shading). But the recovery of shape would be invalid if the shading actually arose from reflectance variation. When a corrugated surface is painted with an albedo texture, the variation in local mean luminance (LM) due to shading is accompanied by a similar modulation in texture amplitude (AM). This is not so for reflectance variation, nor for roughly textured surfaces. We used a haptic matching technique to show that modulations of texture amplitude play a role in the interpretation of shape from shading. Observers were shown plaid stimuli comprising LM and AM combined in-phase (LM+AM) on one oblique and in anti-phase (LM-AM) on the other. Stimuli were presented via a modified ReachIN workstation allowing the co-registration of visual and haptic stimuli. In the first experiment, observers were asked to adjust the phase of a haptic surface, which had the same orientation as the LM+AM combination, until its peak in depth aligned with the visually perceived peak. The resulting alignments were consistent with the use of a lighting-from-above prior. In the second experiment, observers were asked to adjust the amplitude of the haptic surface to match that of the visually perceived surface. Observers chose relatively large amplitude settings when the haptic surface was oriented and phase-aligned with the LM+AM cue. When the haptic surface was aligned with the LM-AM cue, amplitude settings were close to zero. Thus the LM/AM phase relation is a significant visual depth cue, and is used to discriminate between shading and reflectance variations. [Supported by the Engineering and Physical Sciences Research Council, EPSRC].

The role of texture in shape from shading: are humans biased towards seeing relief textures?

When a textured surface is modulated in depth and illuminated, the level of illumination varies across the surface, producing coarse-scale luminance modulations (LM) and amplitude modulation (AM) of the fine-scale texture. If the surface has an albedo texture (reflectance variation) then the LM and AM components are always in-phase, but if the surface has a relief texture the phase relation between LM and AM varies with the direction and nature of the illuminant. We showed observers sinusoidal luminance and amplitude modulations of a binary noise texture, in various phase relationships, in a paired-comparisons design. In the first experiment, the combinations under test were presented in different temporal intervals. Observers indicated which interval contained the more depthy stimulus. LM and AM in-phase were seen as more depthy than LM alone which was in turn more depthy than LM and AM in anti-phase, but the differences were weak. In the second experiment the combinations under test were presented in a single interval on opposite obliques of a plaid pattern. Observers were asked to indicate the more depthy oblique. Observers produced the same depth rankings as before, but now the effects were more robust and significant. Intermediate LM/AM phase relationships were also tested: phase differences less than 90 deg were seen as more depthy than LM-only, while those greater than 90 deg were seen as less depthy. We conjecture that the visual system construes phase offsets between LM and AM as indicating relief texture and thus perceives these combinations as depthy even when their phase relationship is other than zero. However, when different LM/AM pairs are combined in a plaid, the signals on the obliques are unlikely to indicate corrugations of the same texture, and in this case the out-of-phase pairing is seen as flat. [Supported by the Engineering and Physical Sciences Research Council (EPSRC)].

The role of texture amplitude in shape from shading

When a textured surface is modulated in depth and illuminated, parts of the surface receive different levels of illumination; the resulting variations in luminance can be used to infer the shape of the depth modulations-shape from shading. The changes in illumination also produce changes in the amplitude of the texture, although local contrast remains constant. We investigated the role of texture amplitude in supporting shape from shading. If a luminance plaid is added to a binary noise texture (LM), shape from shading produces perception of corrugations in two directions. If the amplitude of the noise is also modulated (AM) such that it is in-phase with one of the luminance sinusoids and out-of-phase with the other, the resulting surface is seen as corrugated in only one directionöthat supported by the in-phase pairing. We confirmed this subjective report experimentally, using a depth-mapping technique. Further, we asked naïve observers to indicate the direction of corrugations in plaids made up of various combinations of LM and AM. LM+AM was seen as having most depth, then LM-only, then LM-AM, and then AM-only. Our results suggest that while LM is required to see depth from shading, its phase relative to any AM component is also important.

The interaction of luminance and texture amplitude in surface depth perception

Previous studies have suggested separate channels for detection of first-order luminance modulations (LM) and second-order modulations of the local amplitude (AM) of a texture. Mixtures of LM and AM with different phase relationships appear very different: in-phase compounds (LM + AM) look like 3-D corrugated surfaces, while out-of-phase compounds (LM - AM) appear flat and/or transparent. This difference may arise because the in-phase compounds are consistent with multiplicative shading, while the out-of-phase compounds are not. We investigated the role of these modulation components in surface depth perception. We used a textured background with thin bars formed by local changes in luminance and/or texture amplitude. These stimuli appear as embossed surfaces with wide and narrow regions. Keeping the AM modulation depth fixed at a suprathreshold level, we determined the amount of luminance contrast required for observers to correctly indicate the width (narrow or wide) of 'raised' regions in the display. Performance (compared to the LM-only case) was facilitated by the presence of AM, but, unexpectedly, performance for LM - AM was as good as for LM + AM. Thus, these results suggest that there is an interaction between first-order and second-order mechanisms during depth perception based on shading cues, but the phase dependence is not yet understood.

The interaction of luminance and texture amplitude in depth perception

Previous studies have suggested separate channels for the detection of first-order luminance (LM) and second-order modulations of the local amplitude (AM) of a texture (Schofield and Georgeson, 1999 Vision Research 39 2697 - 2716; Georgeson and Schofield, 2002 Spatial Vision 16 59). It has also been shown that LM and AM mixtures with different phase relationships are easily separated in identification tasks, and (informally) appear very different with the in-phase compound (LM + AM), producing the most realistic depth percept. We investigated the role of these LM and AM components in depth perception. Stimuli consisted of a noise texture background with thin bars formed as local increments or decrements in luminance and/or noise amplitude. These stimuli appear as embossed surfaces with wide and narrow regions. When luminance and amplitude changes have the same sign and magnitude (LM + AM) the overall modulation is consistent with multiplicative shading, but this is not so when the two modulations have opposite sign (LM - AM). Keeping the AM modulation depth fixed at a suprathreshold level, we determined the amount of luminance contrast required for observers to correctly indicate the width (narrow or wide) of raised regions in the display. Performance (compared to the LM-only case) was facilitated by the presence of AM, but, unexpectedly, performance for LM - AM was even better than for LM + AM. Further tests suggested that this improvement in performance is not due to an increase in the detectability of luminance in the compound stimuli. Thus, contrary to previous findings, these results suggest the possibility of interaction between first-order and second-order mechanisms in depth perception.

Shading and texture:Separate information channels with a common adaptation mechanism?

We outline a scheme for the way in which early vision may handle information about shading (luminance modulation, LM) and texture (contrast modulation, CM). Previous work on the detection of gratings has found no sub-threshold summation, and no cross-adaptation, between LM and CM patterns. This strongly implied separate channels for the detection of LM and CM structure. However, we now report experiments in which adapting to LM (or CM) gratings creates tilt aftereffects of similar magnitude on both LM and CM test gratings, and reduces the perceived strength (modulation depth) of LM and CM gratings to a similar extent. This transfer of aftereffects between LM and CM might suggest a second stage of processing at which LM and CM information is integrated. The nature of this integration, however, is unclear and several simple predictions are not fulfilled. Firstly, one might expect the integration stage to lose identity information about whether the pattern was LM or CM. We show instead that the identity of barely detectable LM and CM patterns is not lost. Secondly, when LM and CM gratings are combined in-phase or out-of-phase we find no evidence for cancellation, nor for 'phase-blindness'. These results suggest that information about LM and CM is not pooled or merged - shading is not confused with texture variation. We suggest that LM and CM signals are carried by separate channels, but they share a common adaptation mechanism that accounts for the almost complete transfer of perceptual aftereffects.

Texture description and segmentation in image processing

Textured regions in images can be defined as those regions containing a signal which has some measure of randomness. This thesis is concerned with the description of homogeneous texture in terms of a signal model and to develop a means of spatially separating regions of differing texture. A signal model is presented which is based on the assumption that a large class of textures can adequately be represented by their Fourier amplitude spectra only, with the phase spectra modelled by a random process. It is shown that, under mild restrictions, the above model leads to a stationary random process. Results indicate that this assumption is valid for those textures lacking significant local structure. A texture segmentation scheme is described which separates textured regions based on the assumption that each texture has a different distribution of signal energy within its amplitude spectrum. A set of bandpass quadrature filters are applied to the original signal and the envelope of the output of each filter taken. The filters are designed to have maximum mutual energy concentration in both the spatial and spatial frequency domains thus providing high spatial and class resolutions. The outputs of these filters are processed using a multi-resolution classifier which applies a clustering algorithm on the data at a low spatial resolution and then performs a boundary estimation operation in which processing is carried out over a range of spatial resolutions. Results demonstrate a high performance, in terms of the classification error, for a range of synthetic and natural textures

Soft hydrogels from nanotubes of poly(ethylene oxide)−tetraphenylalanine conjugates prepared by click chemistry

A new poly(ethylene oxide)-tetraphenylalanine polymer-peptide conjugate has been prepared via a “click” reaction between an alkyne-modified peptide and an azide-terminated PEO oligomer. Self-assembled nanotubes are formed after dialysis of a THF solution of this polymer-peptide conjugate against water. The structure of these nanotubes has been probed by circular dichroism, IR, TEM, and SAXS. From these data, it is apparent that self-assembly involves the formation of antiparallel ß-sheets and p-p-stacking. Nanotubes are formed at concentrations between 2 and 10 mg mL-1. Entanglement between adjacent nanotubes occurs at higher concentrations, resulting in the formation of soft hydrogels. Gel strength increases at higher polymer-peptide conjugate concentration, as expected.

Analytical techniques for the study of soft contact lens spoilation

Soft contact lens wear has become a common phenomenon in recent times. The contact lens when placed in the eye rapidly undergoes change. A film of biological material builds up on and in the lens matrix. The long term wear characteristics of the lens ultimately depend on this process. With time distinct structures made up of biological material have been found to build up on the lens. A fuller understanding of this process and how it relates to the lens chemistry could lead to contact lenses that are better tolerated by the eye. The tear film is a complex biological fluid, it is this fluid that bathes the lens during wear. It is reasonable to suppose that it is material derived from this source that accumulates on the lens. To understand this phenomenon it was decided to investigate the make up and conformation of the protein species that are found on and in the lens. As inter individual variations in tear fluid composition have been found it is important to be able to study the proteins on a single lens. Many of the analytical techniques used in bio research are not suitable for this study because of the lack of sensitivity. Work with poly acrylamide electrophoresis showed the possibility of analyzing the proteins extracted from a single lens. The development of a biotin avidin electro-blot and an enzyme linked aniibody electro-blot, lead to the high sensitivity detection and identification of the proteins present. The extraction of proteins from a lens is always incomplete. A method that analyses the proteins in situ would be a great advancement. Fourier transform infra red microscopy was developed to a point where a thin section of a contact lens could yield information about the proteins present and their conformation. The three dimensional structure of the gross macroscopic structures termed white spots was investigated using confocal laser microscopy.