Approaches for Monitoring Warfighter Blast-related Exposures in Training to Develop Effective Safety Standards.

INTRODUCTION: Traumatic brain injuries are of concern to the sports and military communities because of the age of the participants and costly burden to society. To markedly reduce the impact of traumatic brain injury and its sequela (TBI-S), it is necessary to determine the initial vulnerability of individuals as well as identify new technologies that indicate early signs of TBI-S. MATERIALS AND METHODS: Currently, diverse methods have been used by the authors and others in laboratory settings to reveal early signs of persistent TBI-S including simulation modeling of the effect of rapid deceleration on the deviatoric strain (shear force) imposed on specific brain regions, auditory evoked potential (AEP) measurements to determine injury to the auditory cortex optokinetic nystagmus (OKN) measures sensitive to vestibular trauma, and optical coherence tomography (OCT) measures that reveal changes in central visual function obtained noninvasively by examination of the retina. RESULTS: Simulation studies provided technical information on maximal deviatoric strain at the base of the sulci and interface of gray and white matter consistent with results from neuropathology and from magnetic resonance imaging. The AEP and OKN reveal measurable injury to similar regions below the Sylvian fissure including auditory cortex and midbrain, and the OCT reveals changes to the retina consistent with forceful deceleration effects. CONCLUSIONS: The studies and results are consistent with prior work demonstrating that noninvasive tests may be sensitive to the presence of TBI-S, potentially in the training field as advances in the portability of test instruments are underway. When combined with baseline data gathered from individuals in quantitative form, key variances can emerge. Therefore, it is hypothesized that AEP, OKN, and OCT, taken together, may yield faster objective and quantitative neurophysiological measures serving as a "signature" of neural injury and more indicative of potentially persistent TBI-S-recommending larger scale longitudinal studies.

Scattering Angle Resolved Optical Coherence Tomography Detects Early Changes in 3xTg Alzheimer's Disease Mouse Model.

Purpose: Clinical intensity-based optical coherence tomographic retinal imaging is unable to resolve some of the earliest changes to Alzheimer's disease (AD) neurons. The aim of this pilot study was to demonstrate that scattering-angle-resolved optical coherence tomography (SAR-OCT), which is sensitive to changes in light scattering angle, is a candidate retinal imaging modality for early AD detection. SAR-OCT signal data may be sensitive to changes in intracellular constituent morphology that are not detectable with conventional OCT. Methods: In this cross-sectional study, retinas of a triple transgenic mouse model of AD (3xTg-AD) were imaged alongside age-matched control mice (C57BL/6J) using SAR-OCT. A total of 32 mice (12 control, 20 3xTg-Ad) at four ages (10, 20, 30, and 45 weeks) were included in this cross-sectional study, and three retinal feature sets (scattering, thickness, and angiography) were examined between the disease and control groups. Results: AD mice had significantly increased scattering diversity (lower SAR-OCT C parameter) at the earliest imaging time (10 weeks). Differences in the C parameter between AD and control mice were diminished at later times when both groups showed increased scattering diversity. AD mice have reduced retinal thickness compared to controls, particularly in central regions and superficial layers. No differences in vascular density or fractional blood volume between groups were detected. Conclusions: SAR-OCT is sensitive to scattering angle changes in a 3xTg-AD mouse model and could provide early-stage biomarkers for neurodegenerative diseases such as AD. Translational Relevance: Clinical OCT systems may be modified to record SAR-OCT images for non-invasive retinal diagnostic imaging of patients with neurodegenerative diseases such as AD.

Scattering-Angle-Resolved Optical Coherence Tomography of a Hypoxic Mouse Retina Model.

Several studies have noted a correlation between retinal degeneration and traumatic encephalopathy (TE) making the retina a leading candidate for detection and assessment. Scattering-angle-resolved optical coherence tomography (SAR-OCT) is a candidate imaging modality to detect sub-resolution changes in retinal microstructure. SAR-OCT images of murine retinas that experience a hypoxic insult-euthanasia by isoflurane overdose-are presented. A total of 4 SAR-OCT measurement parameters are reported in 6 longitudinal experiments: blood flow volume fraction, total retinal thickness, reflectance index, and scattering angle. As each mouse expires, blood flow volume fraction decreases, total retinal thickness increases, reflectance index decreases, and scattering angle diversity increases. Contribution of the retinal vasculature to scattering angle diversity is discussed. Results of this study suggest the utility of SAR-OCT to measure TE using scattering angle diversity contrast in the retina.

Fourier optics analysis of phase-mask-based path-length-multiplexed optical coherence tomography.

Optical coherence tomography (OCT) is an imaging technique that constructs a depth-resolved image by measuring the optical path-length difference between broadband light backscattered from a sample and a reference surface. For many OCT sample arm optical configurations, sample illumination and backscattered light detection share a common path. When a phase mask is placed in the sample path, features in the detected signal are observed, which suggests that an analysis of a generic common path OCT imaging system is warranted. In this study, we present a Fourier optics analysis using a Fresnel diffraction approximation of an OCT system with a path-length-multiplexing element (PME) inserted in the sample arm optics. The analysis may be generalized for most phase-mask-based OCT systems. A radial-angle-diverse PME is analyzed in detail, and the point spread function, coherent transfer function, sensitivity of backscattering angular diversity detection, and signal formation in terms of sample spatial frequency are simulated and discussed. The analysis reveals important imaging features and application limitations of OCT imaging systems with a phase mask in the sample path optics.

Degradation in the degree of polarization in human retinal nerve fiber layer.

Using a fiber-based swept-source (SS) polarization-sensitive optical coherence tomography (PS-OCT) system, we investigate the degree of polarization (DOP) of light backscattered from the retinal nerve fiber layer (RNFL) in normal human subjects. Algorithms for processing data were developed to analyze the deviation in phase retardation and intensity of backscattered light in directions parallel and perpendicular to the nerve fiber axis (fast and slow axes of RNFL). Considering superior, inferior, and nasal quadrants, we observe the strongest degradation in the DOP with increasing RNFL depth in the temporal quadrant. Retinal ganglion cell axons in normal human subjects are known to have the smallest diameter in the temporal quadrant, and the greater degradation observed in the DOP suggests that higher polarimetric noise may be associated with neural structure in the temporal RNFL. The association between depth degradation in the DOP and RNFL structural properties may broaden the utility of PS-OCT as a functional imaging technique.

Retinal nerve fiber layer reflectance for early glaucoma diagnosis.

PURPOSE: Compare performance of normalized reflectance index (NRI) and retinal nerve fiber layer thickness (RNFLT) parameters determined from optical coherence tomography (OCT) images for glaucoma and glaucoma suspect diagnosis. METHODS: Seventy-five eyes from 71 human subjects were studied: 33 controls, 24 glaucomatous, and 18 glaucoma-suspects. RNFLT and NRI maps were measured using 2 custom-built OCT systems and the commercial instrument RTVue. Using area under the receiver operating characteristic curve, RNFLT and NRI measured in 7 RNFL locations were analyzed to distinguish between control, glaucomatous, and glaucoma-suspect eyes. RESULTS: The mean NRI of the control group was significantly larger than the means of glaucomatous and glaucoma-suspect groups in most RNFL locations for all 3 OCT systems (P<0.05 for all comparisons). NRI performs significantly better than RNFLT at distinguishing between glaucoma-suspect and control eyes using RTVue OCT (P=0.008). The performances of NRI and RNFLT for classifying glaucoma-suspect versus control eyes were statistically indistinguishable for PS-OCT-EIA (P=0.101) and PS-OCT-DEC (P=0.227). The performances of NRI and RNFLT for classifying glaucomatous versus control eyes were statistically indistinguishable (PS-OCT-EIA: P=0.379; PS-OCT-DEC: P=0.338; RTVue OCT: P=0.877). CONCLUSIONS: NRI is a promising measure for distinguishing between glaucoma-suspect and control eyes and may indicate disease in the preperimetric stage. Results of this pilot clinical study warrant a larger study to confirm the diagnostic power of NRI for diagnosing preperimetric glaucoma.

Micro-patterned drug delivery device for light-activated drug release.

BACKGROUND: The primary goal of this study was the fabrication, long-term stability, and measured release of a marker dye from a micro-patterned drug delivery device using (i) mechanical puncture and (ii) photodisruption with an ophthalmic Nd:YAG laser. MATERIALS AND METHODS: A drug delivery device was made from a transparent bio-compatible polymer. The device consisted of two 2.6 mm diameter reservoirs containing 10% Na fluorescein dye. The device was implanted in the rabbit's eye (n = 2) with the cap of the device facing toward the exterior of the eye. Once the animals recovered from the implant surgery, 100% anhydrous glycerol was topically applied to the eye at the implantation site to decrease light scattering in the conjunctiva and sclera. The dye was released from one of the reservoir either using a 28 G ½ needle or an ophthalmic Q-switched Nd:YAG laser. A fluorescence spectrophotometer (FS) with fiber optic probe was used to measure the half-life of the dye in the eye. Measurements of fluorescence intensity were collected until the measurements return to baseline and histology was done on the tissue surrounded the device. RESULTS: None of the devices leaked of 10% Na fluorescein dye after implant. The ablation threshold of the drug delivery device was between 6 and 10 mJ to create 100-500 µm holes. The half-life measurement of the dye was found to be 13 days at the vitreous chamber after measuring the fluorescence intensity through the dilated cornea. Histology study showed minimal immune and foreign body response such as mild inflammation. CONCLUSION: This study established that the drug delivery device seemed to elicit minimal inflammatory response and retained its fluidic content until it was released with relatively longer retention time (half-life). Thus, similar device could be used for controlled release of drugs for certain ocular diseases.

Changes in morphology and optical properties of sclera and choroidal layers due to hyperosmotic agent.

Light scattering in the normally white sclera prevents diagnostic imaging or delivery of a focused laser beam to a target in the underlying choroid layer. In this study, we examine optical clearing of the sclera and changes in blood flow resulting from the application of glycerol to the sclera of rabbits. Recovery dynamics are monitored after the application of saline. The speed of clearing for injection delivery is compared to the direct application of glycerol through an incision in the conjunctiva. Although, the same volume of glycerol was applied, the sclera cleared much faster (5 to 10 s) with the topical application of glycerol compared to the injection method (3 min). In addition, the direct topical application of glycerol spreads over a larger area in the sclera than the latter method. A diffuse optical spectroscopy system provided spectral analysis of the remitted light every two minutes during clearing and rehydration. Comparison of measurements to those obtained from phantoms with various absorption and scattering properties provided estimates of the absorption coefficient and reduced scattering coefficient of rabbit eye tissue.

Birefringence measurement of the retinal nerve fiber layer by swept source polarization sensitive optical coherence tomography.

A Swept Source Polarization-Sensitive Optical Coherence Tomography (SS-PS-OCT) instrument has been designed, constructed, and verified to provide high sensitivity depth-resolved birefringence and phase retardation measurements of the retinal nerve fiber layer. The swept-source laser had a center wavelength of 1059 nm, a full-width-half-max spectral bandwidth of 58 nm and an A-line scan rate of 34 KHz. Power incident on the cornea was 440 µW and measured axial resolution was 17 µm in air. A multiple polarization state nonlinear fitting algorithm was used to measure retinal birefringence with low uncertainty. Maps of RNFL phase retardation in a subject measured with SS-PS-OCT compare well with those generated using a commercial scanning laser polarimetry instrument. Peak-to-valley variation of RNFL birefringence given here is less than values previously reported at 840nm.

Variation of fluorescence in tissue with temperature.

BACKGROUND AND OBJECTIVE: Previous studies demonstrated a decrease in fluorescence intensity as tissue temperature increased. In vitro samples were increased from room temperature and in vivo canine liver from body temperature. This study investigated variations in fluorescence intensity with temperatures starting at 14°C and compared in vivo and in vitro results for consistency. STUDY DESIGN/MATERIAL AND METHODS: A fiber optic-based noninvasive system was used to characterize the temperature effect on tissue fluorescence in hamster dorsal skin in vivo, and in sclera and cornea of enucleated pig eyes in vitro. As tissue was allowed to progress through the temperature range of 14-42°C, the spectra of auto-fluorescence with respect to temperature was sampled every 1-2 minutes. A pulsed nitrogen laser was used to excite fluorescence through a fiber optic probe with a source-detector aperture separation of 370 µm. RESULTS: Fluorescence intensity decreased as temperature increased from 14 to 42°C in a phantom containing Rhodamine B dye. Results from both in vivo and in vitro tissue followed the same trend of decreasing intensity as tissue temperature increased from 14°C. Spectral intensity lineshape changed around 450 nm due to absorption from tissue. CONCLUSION: Cooling a tissue increased fluorescence intensity of skin in vivo, in all experiments. In vitro results were consistent with in vivo measurements.

Perfusion in hamster skin treated with glycerol.

BACKGROUND AND OBJECTIVE: The objective of this article is to quantify the effect of hyper-osmotic agent (glycerol) on blood velocity in hamster skin blood vessels measured with a dynamic imaging technique, laser speckle contrast imaging (LSCI). STUDY DESIGN/MATERIALS AND METHODS: In this study a dorsal skin-flap window was implanted on the hamster skin. The hyper-osmotic drug, that is, glycerol was delivered to the skin through the open dermal end of the window model. A two-dimensional map of blood flow of skin blood vessels was obtained from the speckle contrast (SC) images. RESULTS: Preliminary studies demonstrated that hyper-osmotic agents such as glycerol not only make tissue temporarily transparent, but also reduce blood flow. The blood perfusion was measured every 3 minutes for 36-66 minutes after diffusion of anhydrous glycerol. Blood flow in small capillaries was found to be reduced significantly within 3-9 minutes. Blood flow in larger blood vessels (i.e., all arteries and veins) decreased over time and some veins had significantly reduced blood flow within 36 minutes. At 24 hours, there was a further reduction in capillary blood perfusion whereas larger blood vessels regained flow compared to an hour after initial application of glycerol. CONCLUSION: Blood flow velocity and vessel diameter of the micro-vasculatures of hamster skin were reduced by the application of 100% anhydrous glycerol. At 24 hours, capillary perfusion remained depressed.