Polarized Light Imaging Research Articles

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Next article FreeAbout the CoverPDFPDF PLUSFull Text Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinked InRedditEmailQR Code SectionsMoreA defining feature of echinoderms is their five-fold symmetry, which arises during the metamorphosis of a bilaterally symmetrical larva. During echinoderm metamorphosis, the coelomic cavities undergo an elaborate transformation from early mesodermal pouches to a complex system of tubes and rings that make up the numerous internal components of the animal. In this issue (pp. 283–298), H. C. Sweet and several undergraduate research students (G. Azriel, N. Jaff, J. Moser, T. A. Riola, C. Ideman, M. Barton, J. Nelson, and M. M. Lenhart) demonstrate the origin of the coelomic structures and their morphogenesis in the vitellaria larva of the brittle star Ophioplocus esmarki. Many of the structures first form when the archenteron becomes subdivided into an unpaired axocoel, hydrocoel, somatocoel, and stomach rudiment. The right somatocoel and part of the left somatocoel originate as bilateral invaginations of the larval ectoderm. This supports the findings of others that ectodermal invagination of the somatocoels is an unusual feature of brittle star development. Confocal microscopy and 3D modeling were used to document the later morphogenesis of the coelomic cavities as they form juvenile structures with five-fold symmetry.The cover image demonstrates the complex structure of the vitellaria larva. The image shows the color-coded coelomic cavities superimposed with an image of the skeleton and an outline of body of the vitellaria larva. At this stage, the outer ectoderm has ciliary bands on raised ridges that are used for swimming (grayscale outline), but most of the internal structures are patterned in five-fold symmetry with the axial complex near the anterior end (top). Coelomic structures include the water vascular system derived from the hydrocoel (yellow), hypogastric coelom derived from the left somatocoel (purple), genital coelom (peach) derived from the left somatocoel, epigastric coelom derived from the right somatocoel (magenta), pericardial coelom (green), axial coelom (orange), and pore canal (red). The juvenile skeletal plates form adjacent to the coelomic structures and the overlying ectoderm. The image was generated with confocal microscopy, polarized light imaging, and image colorizing and processing in GIMP and Fiji.Credits: Photo, Hyla C. Sweet; cover design, Olivia Kinker. Next article DetailsFiguresReferencesCited by The Biological Bulletin Volume 243, Number 3December 2022 Published in association with the Marine Biological Laboratory Article DOIhttps://doi.org/10.1086/724434 © 2022 The University of Chicago. All rights reserved.PDF download Crossref reports no articles citing this article.

Computational Portable Microscopes for Point-of-Care-Test and Tele-Diagnosis.

In bio-medical mobile workstations, e.g., the prevention of epidemic viruses/bacteria, outdoor field medical treatment and bio-chemical pollution monitoring, the conventional bench-top microscopic imaging equipment is limited. The comprehensive multi-mode (bright/dark field imaging, fluorescence excitation imaging, polarized light imaging, and differential interference microscopy imaging, etc.) biomedical microscopy imaging systems are generally large in size and expensive. They also require professional operation, which means high labor-cost, money-cost and time-cost. These characteristics prevent them from being applied in bio-medical mobile workstations. The bio-medical mobile workstations need microscopy systems which are inexpensive and able to handle fast, timely and large-scale deployment. The development of lightweight, low-cost and portable microscopic imaging devices can meet these demands. Presently, for the increasing needs of point-of-care-test and tele-diagnosis, high-performance computational portable microscopes are widely developed. Bluetooth modules, WLAN modules and 3G/4G/5G modules generally feature very small sizes and low prices. And industrial imaging lens, microscopy objective lens, and CMOS/CCD photoelectric image sensors are also available in small sizes and at low prices. Here we review and discuss these typical computational, portable and low-cost microscopes by refined specifications and schematics, from the aspect of optics, electronic, algorithms principle and typical bio-medical applications.

Quantative MRI predicts tendon mechanical behavior, collagen composition, and organization.

Quantitative magnetic resonance imaging (qMRI) measures have provided insights into the composition, quality, and structure-function of musculoskeletal tissues. Low signal-to-noise ratio has limited application to tendon. Advances in scanning sequences and sample positioning have improved signal from tendon allowing for evaluation of structure and function. The purpose of this study was to elucidate relationships between tendon qMRI metrics (T1, T2, T1ρand diffusion tensor imaging [DTI] metrics) with tendon tissue mechanics, collagen concentration and organization. Sixteen human Achilles tendon specimens were collected, imaged with qMRI, and subjected to mechanical testing with quantitative polarized light imaging. T2 values were related to tendon mechanics [peak stress (rsp = 0.51, p = 0.044), equilibrium stress (rsp = 0.54, p = 0.033), percent relaxation (rsp = -0.55, p = 0.027), hysteresis (rsp = -0.64, p = 0.007), linear modulus (rsp = 0.67, p = 0.009)]. T1ρhad a statistically significant relationship with percent relaxation (r = 0.50, p = 0.048). Collagen content was significantly related to DTI measures (range of r = 0.56-0.62). T2 values from a single slice of the midportion of human Achilles tendons were strongest predictors of tendon tensile mechanical metrics. DTI diffusivity indices (mean diffusivity, axial diffusivity, radial diffusivity) were strongly correlated with collagen content. These findings build on a growing body of literature supporting the feasibility of qMRI to characterize tendon tissue and noninvasively measure tendon structure and function. Statement of Clinical Significance: Quantitative MRI can be applied to characterize tendon tissue and is a noninvasive measure that relates to tendon composition and mechanical behavior.

Backscattering Mueller Matrix polarimetry on whole brain specimens shows promise for minimally invasive mapping of microstructural orientation features

Understanding microscale physiology and microstructural cellular features of the brain is key to understanding mechanisms of neurodegenerative diseases and injury, as well as prominent changes undergone in development and aging. Non-invasive imaging modalities sensitive to the microscale, especially diffusion magnetic resonance imaging (dMRI), are promising for mapping of cellular microstructure of brain tissues; however, there is a need for robust validation techniques to verify and improve the biological accuracy of information derived. Recent advances in dMRI have moved toward probing of the more complex grey matter architecture, challenging current validation techniques, which are largely based on ex vivo staining and microscopy focusing on white matter. Polarized light imaging (PLI) has been shown to be successful for high resolution, direct, microstructural imaging and has been applied to dMRI validation with clear advantages over staining and microscopy techniques. Conventionally, PLI is applied to thin, sectioned samples in transmission mode, but PLI has also been extended to operate in reflectance mode to bridge the gap toward in vivo measurements of the brain. In this report we investigate the use of backscattering Mueller Matrix polarimetry to characterize the microstructural content of intact ferret brain specimens. The results show that backscattering polarimetry can probe white matter fiber coherence and fiber orientation, and show promise for probing grey matter microstructure. Ultimately, this motivates further study to fully understand how best to implement backscattering polarimetry for in vivo microstructural imaging of the brain.

Overview of the microanatomy of the human brainstem in relation to the safe entry zones.

The primary objective of this anatomical study was to apply innovative imaging techniques to increase understanding of the microanatomical structures of the brainstem related to safe entry zones. The authors hypothesized that such a high-detail overview would enhance neurosurgeons' abilities to approach and define anatomical safe entry zones for use with microsurgical resection techniques for intrinsic brainstem lesions. The brainstems of 13 cadavers were studied with polarized light imaging (PLI) and 11.7-T MRI. The brainstem was divided into 3 compartments-mesencephalon, pons, and medulla-for evaluation with MRI. Tissue was further sectioned to 100 μm with a microtome. MATLAB was used for further data processing. Segmentation of the internal structures of the brainstem was performed with the BigBrain database. Thirteen entry zones were reported and assessed for their safety, including the anterior mesencephalic zone, lateral mesencephalic sulcus, interpeduncular zone, intercollicular region, supratrigeminal zone, peritrigeminal zone, lateral pontine zone, median sulcus, infracollicular zone, supracollicular zone, olivary zone, lateral medullary zone, and anterolateral sulcus. The microanatomy, safety, and approaches are discussed. PLI and 11.7-T MRI data show that a neurosurgeon possibly does not need to consider the microanatomical structures that would not be visible on conventional MRI and tractography when entering the mentioned safe entry zones. However, the detailed anatomical images may help neurosurgeons increase their understanding of the internal architecture of the human brainstem, which in turn could lead to safer neurosurgical intervention.

Discrimination and quantification of scar tissue by Mueller matrix imaging with machine learning

Scarring is one of the biggest areas of unmet need in the long-term success of glaucoma filtration surgery. Quantitative evaluation of the scar tissue and the post-operative structure with micron scale resolution facilitates development of anti-fibrosis techniques. However, the distinguishment of conjunctiva, sclera and the scar tissue in the surgical area still relies on pathologists’ experience. Since polarized light imaging is sensitive to anisotropic properties of the media, it is ideal for discrimination of scar in the subconjunctival and episcleral area by characterizing small differences between proportion, organization and the orientation of the fibers. In this paper, we defined the conjunctiva, sclera, and the scar tissue as three target tissues after glaucoma filtration surgery and obtained their polarization characteristics from the tissue sections by a Mueller matrix microscope. Discrimination score based on parameters derived from Mueller matrix and machine learning was calculated and tested as a diagnostic index. As a result, the discrimination score of three target tissues showed significant difference between each other ([Formula: see text]). The visualization of the discrimination results showed significant contrast between target tissues. This study proved that Mueller matrix imaging is effective in ocular scar discrimination and paves the way for its application on other forms of ocular fibrosis as a substitute or supplementary for clinical practice.

Fast data-driven computation and intuitive visualization of fiber orientation uncertainty in 3D-polarized light imaging

In recent years, the microscopy technology referred to as Polarized Light Imaging (3D-PLI) has successfully been established to study the brain’s nerve fiber architecture at the micrometer scale. The myelinated axons of the nervous tissue introduce optical birefringence that can be used to contrast nerve fibers and their tracts from each other. Beyond the generation of contrast, 3D-PLI renders the estimation of local fiber orientations possible. To do so, unstained histological brain sections of 70 μm thickness cut at a cryo-microtome were scanned in a polarimetric setup using rotating polarizing filter elements while keeping the sample unmoved. To address the fundamental question of brain connectivity, i. e., revealing the detailed organizational principles of the brain’s intricate neural networks, the tracing of fiber structures across volumes has to be performed at the microscale. This requires a sound basis for describing the in-plane and out-of-plane orientations of each potential fiber (axis) in each voxel, including information about the confidence level (uncertainty) of the orientation estimates. By this means, complex fiber constellations, e. g., at the white matter to gray matter transition zones or brain regions with low myelination (i. e., low birefringence signal), as can be found in the cerebral cortex, become quantifiable in a reliable manner. Unfortunately, this uncertainty information comes with the high computational price of their underlying Monte-Carlo sampling methods and the lack of a proper visualization. In the presented work, we propose a supervised machine learning approach to estimate the uncertainty of the inferred model parameters. It is shown that the parameter uncertainties strongly correlate with simple, physically explainable features derived from the signal strength. After fitting these correlations using a small sub-sample of the data, the uncertainties can be predicted for the remaining data set with high precision. This reduces the required computation time by more than two orders of magnitude. Additionally, a new visualization of the derived three-dimensional nerve fiber information, including the orientation uncertainty based on ellipsoids, is introduced. This technique makes the derived orientation uncertainty information visually interpretable.

Single shot quantitative polarized light imaging system for rapid planar biaxial testing of soft tissues.

Quantitative Polarized Light Imaging (QPLI) is an established technique used to compute the orientation of collagen fibers based on their birefringence. QPLI systems typically require rotating linear polarizers to obtain sufficient data to estimate orientation, which limits acquisition speeds and is not ideal for its application to mechanical testing. In this paper, we present a QPLI system designed with no moving parts; a single shot technique which is ideal to characterize collagen fiber orientation and kinematics during mechanical testing. Our single shot QPLI system (ssQPLI) sorts polarized light into four linear polarization states that are collected simultaneously by four cameras. The ssQPLI system was validated using samples with known orientation and retardation, and we demonstrate its use with planar biaxial testing of mouse skin. The ssQPLI system was accurate with a mean orientation error of 1.35° ± 1.58°. Skin samples were tested with multiple loading protocols and in each case the mean orientation of the collagen network reoriented to align in the direction of primary loading as expected. In summary, the ssQPLI system is effective at quantifying collagen fiber organization, and, when combined with mechanical testing, can rapidly provide pixel-wise measures of fiber orientation during biaxial loading.

Single-kernel classification of deoxynivalenol and zearalenone contaminated maize based on visible light imaging under ultraviolet light excitation combined with polarized light imaging

Deoxynivalenol and zearalenone contamination in maize poses a threat to food safety. Thus, the development of a cost-effective and non-destructive method to classify deoxynivalenol and zearalenone contaminated maize is an important research issue. Natural deoxynivalenol and zearalenone contaminated maize were randomly selected for this experiment. Visible light imaging under ultraviolet light excitation combined with polarized light imaging was used to extract texture and color features from the endosperm and germ sides. And six classification models were established for the single-kernel classification of 1035 contaminated maize. The results indicated that the optimal detection surface was the endosperm side, and the optimal model was SVM with a classification accuracy of 88.1%. Then, local feature analysis of images and mycotoxin concentrations verification experiment were used to reduce secondary contamination and further evaluate the performance of the model, respectively. This method can be verified to be effective in identifying three deoxynivalenol and zearalenone contamination levels of maize, and can also reduce the costs of test stands.

The Application of Light-Assisted Drying to the Thermal Stabilization of Nucleic Acid Nanoparticles.

Background: Cold-chain storage can be challenging and expensive for the transportation and storage of biologics, especially in low-resource settings. Nucleic acid nanoparticles (NANPs) are an example of new biological products that require refrigerated storage. Light-assisted drying (LAD) is a new processing technique to prepare biologics for anhydrous storage in a trehalose amorphous solid matrix at ambient temperatures. In this study, LAD was used to thermally stabilize four types of NANPs with differing structures and melting temperatures. Methods: Small volume samples (10 μL) containing NANPs were irradiated with a 1064 nm laser to speed the evaporation of water and create an amorphous trehalose preservation matrix. Samples were then stored for 1 month at 4°C or 20°C. A FLIR C655 mid-IR camera was used to record the temperature of samples during processing. The trehalose matrix was characterized using polarized light imaging (PLI) to determine if crystallization occurred during processing or storage. Damage to LAD-processed NANPs was assessed after processing and storage using gel electrophoresis. Results: Based on the end moisture content (EMC) as a function time and the thermal histories of samples, a LAD processing time of 30 min is sufficient to achieve low EMCs for the 10 μL samples used in this study. PLI demonstrates that the trehalose matrix was resistant to crystallization during processing and after storage at 4°C and at room temperature. The native-polyacrylamide gel electrophoresis results for DNA cubes, RNA cubes, and RNA rings indicate that the main structures of these NANPs were not damaged significantly after LAD processing and being stored at 4°C or at room temperature for 1 month. Conclusions: These preliminary studies indicate that LAD processing can stabilize NANPs for dry-state storage at room temperature, providing an alternative to refrigerated storage for these nanomedicine products.

Ex vivo experimental characterizations for understanding the interrelationship between tissue mechanics and collagen microstructure of porcine mitral valve leaflets

Unidirectional blood flow in the left side of the heart is regulated by the mitral valve. To better understand the mitral valve function, researchers have examined the structural and mechanical properties of the mitral valve leaflets; however, limitations of the previous studies include the use of mechanics- and structure-altering tissue modifications (e.g., optical clearing) that limit the ability to quantify the unique load-dependent reorientation and realignment of the collagen fibers as well as their interrelation with the valve tissue mechanics. Herein, we aimed to circumvent these limitations by utilizing an integrated polarized-light imaging and biaxial testing system for understanding the mechanics–microstructure interrelationship for porcine mitral valve leaflets. We further performed constitutive modeling and evaluated the accuracy of the affine fiber kinematics theory. From the tissue mechanics perspective, the posterior leaflet was more extensible in the radial direction than the anterior leaflet (14.2% difference in radial tissue stretch), while exhibiting smaller collagen and elastin moduli based on the determined constitutive model parameters. From the collagen microstructure’s standpoint, the posterior leaflet had smaller increases in optical anisotropy (closely related to the degree of fiber alignment) than the anterior leaflet (32.8±7.7% vs. 50.0±19.7%). Further, the leaflets were found to possess two distinct fiber families – one family oriented along the circumferential tissue direction, and another more disperse family with a 30°–40° offset from the first fiber family. Finally, affine fiber kinematics consistently underpredicted the collagen fiber reorientations Overall, this study improved our understanding of the mitral valve leaflets that is essential for facilitating tissue-emulated valve replacement and cardiac valve modeling frameworks.

Linking the region-specific tissue microstructure to the biaxial mechanical properties of the porcine left anterior descending artery

Coronary atherosclerosis is the main cause of death worldwide. Advancing the understanding of coronary microstructure-based mechanics is fundamental for the development of therapeutic tools and surgical procedures. Although the passive biaxial properties of the coronary arteries have been extensively explored, their regional differences and the relationship between tissue microstructure and mechanics have not been fully characterized. In this study, we characterized the passive biaxial mechanical properties and microstructural properties of the proximal, medial, and distal regions of the porcine left anterior descending artery (LADA). We also attempted to relate the biaxial stress-stretch response of the LADA and its respective birefringent responses to the polarized light for obtaining information about the load-dependent microstructural variations. We found that the LADA extensibility is reduced in the proximal-to-distal direction and that the medial region exhibits more heterogeneous mechanical behavior than the other two regions. We have also observed highly dynamic microstructural behavior where fiber families realign themselves depending on loading. In addition, we found that the microstructure of the distal region exhibited highly aligned fibers along the longitudinal axis of the artery. To verify this microstructural feature, we imaged the LADA specimens with multi-photon microscopy and observed that the adventitia microstructure transitioned from a random fiber network in the proximal region to highly aligned fibers in the distal region. Our findings could offer new perspectives for understanding coronary mechanics and aid in the development of tissue-engineered vascular grafts, which are currently limited due to their mismatch with native tissue in terms of mechanical properties and microstructural features. Statement of significanceThe tissue biomechanics of coronary arteries is fundamental for the development of revascularization techniques such as coronary artery bypass. These therapeutics require a deep understanding of arterial mechanics, microstructure, and mechanobiology to prevent graft failure and reoperation. The present study characterizes the unique regional mechanical and microstructural properties of the porcine left anterior descending artery using biaxial testing, polarized-light imaging, and confocal microscopy. This comprehensive characterization provides an improved understanding of the collagen/elastin architecture in response to mechanical loads using a region-specific approach. The unique tissue properties obtained from this study will provide guidance for the selection of anastomotic sites in coronary artery bypass grafting and for the design of tissue-engineered vascular grafts.

Mechanical and Microstructural Properties of Meniscus Roots Vary by Location

Background: Despite the growing awareness of the clinical significance of meniscus root tears, there are relatively limited biomechanical and microstructural data available on native meniscus roots that could improve our understanding of why they are injured and how to best treat them. Purpose/Hypothesis: The purpose of the study was to measure the material and microstructural properties of meniscus roots using mechanical testing and quantitative polarized light imaging. The hypothesis was that these properties vary by location (medial vs lateral, anterior vs posterior) and by specific root (anteromedial vs anterolateral, posteromedial vs posterolateral). Study Design: Descriptive laboratory study. Methods: Anterior and posterior meniscus roots of the medial and lateral meniscus were isolated from 22 cadavers (10 female, 12 male; mean ± SD age, 47.1 ± 5.1 years) and loaded in uniaxial tension. Quantitative polarized light imaging was used to measure collagen fiber organization and realignment under load. Samples were subjected to preconditioning, stress-relaxation, and a ramp to failure. Time-dependent relaxation behavior was quantified. Modulus values were computed in the toe and linear regions of the stress-strain curves. The degree of linear polarization (DoLP) and angle of polarization—measures of the strength and direction of collagen alignment, respectively—were calculated during the stress-relaxation test and at specific strain values throughout the ramp to failure (zero, transition, and linear strain). Results: Anterior roots had larger moduli than posterior roots in the toe (P = .007) and linear (P < .0001) regions and larger average DoLP values at all points of the ramp to failure (zero, P = .016; transition, P = .004; linear, P = .002). Posterior roots had larger values across all regions in terms of standard deviation angle of polarization (P < .001). Lateral roots had greater modulus values versus medial roots in the toe (P = .027) and linear (P = .014) regions. Across all strain points, posterolateral roots had smaller mean DoLP values than posteromedial roots. Conclusion: Posterior meniscus roots have smaller modulus values and more disorganized collagen alignment at all strain levels when compared with anterior roots. Posterolateral roots have lower strength of collagen alignment versus posteromedial roots. Clinical Relevance: These data findings may explain at least in part the relative paucity of anterior meniscus root tears and the predominance of traumatic posterolateral roots tears as compared with degenerative posteromedial root tears.

Green Topochemical Esterification Effects on the Supramolecular Structure of Chitin Nanocrystals: Implications for Highly Stable Pickering Emulsions.

In nature, chitin is organized in hierarchical structures composed of nanoscale building blocks that show outstanding mechanical and optical properties attractive for nanomaterial design. For applications that benefit from a maximized interface such as nanocomposites and Pickering emulsions, individualized chitin nanocrystals (ChNCs) are of interest. However, when extracted in water suspension, their individualization is affected by ChNC self-assembly, requiring a large amount of water (above 90%) for ChNC transport and stock, which limits their widespread use. To master their individualization upon drying and after regeneration, we herein report a waterborne topochemical one-pot acid hydrolysis/Fischer esterification to extract ChNCs from chitin and simultaneously decorate their surface with lactate or butyrate moieties. Controlled reaction conditions were designed to obtain nanocrystals of a comparable aspect ratio of about 30 and a degree of modification of about 30% of the ChNC surface, under the rationale to assess the only effect of the topochemistry on ChNC supramolecular organization. The rheological analysis coupled with polarized light imaging shows how the nematic structuring is hindered by both surface ester moieties. The increased viscosity and elasticity of the modified ChNC colloids indicate a gel-like phase, where typical ChNC clusters of liquid crystalline phases are disrupted. Pickering emulsions have been prepared from lyophilized nanocrystals as a proof of concept. Our results demonstrate that only the emulsions stabilized by the modified ChNCs have excellent stability over time, highlighting that their individualization can be regenerated from the dry state.

Connecting macroscopic diffusion metrics of cardiac diffusion tensor imaging and microscopic myocardial structures based on simulation.

To investigate the relationship between microscopic myocardial structures and macroscopic measurements of diffusion tensor imaging (DTI), we proposed a cardiac DTI simulation method using the Bloch equationand the Monte Carlo random walk in a realistic myocardium model reconstructed from polarized light imaging (PLI) data of the entire human heart. To obtain a realistic simulation, with the constraints of prior knowledge pertaining to the maturational change of the myocardium structure, appropriate microstructure modeling parameters were iteratively determined by matching DTI simulations and real acquisitions of the same hearts in terms of helix angle, fractional anisotropy (FA) and mean diffusivity (MD) maps. Once a realistic simulation was obtained, we varied the extra-cellular volume (ECV) ratio, myocyte orientation heterogeneity and myocyte size, and explored the effects of microscopic changes in tissue structure on macroscopic diffusion metrics. The experimental results demonstrated the feasibility of simulating the DTI of the whole heart using PLI measurements. When varying ECV from 15% to 55%, mean FA decreased from 0.55 to 0.26, axial diffusivity increased by 0.6 μm2/ms, and radial diffusivity increased by 0.7 μm2/ms. When orientation heterogeneity was varied from 0 to 20∘, mean FA decreased from 0.4 to 0.3, axial diffusivity decreased by 0.08 μm2/ms, and radial diffusivity increased by 0.03 μm2/ms. When mean diameter of myocytes was varied from 6 μm to 10 μm, FA decreased from 0.67 to 0.46, axial and radial diffusivities increased by 0.05 and 0.2 μm2/ms, respectively.

Additional fiber orientations in the sagittal stratum—noise or anatomical fine structure?

The sagittal stratum is a prominent and macroscopically clearly visible white-matter structure within occipital and parietal lobes with a highly organized structure of parallel fibers running in rostro-caudal direction. Apart from the major tract running through, i.e., the optic radiation, the source and arrangement of other fibers within the sagittal stratum is only partially understood. Recent diffusion imaging studies in-vivo suggest additional minor fiber directions, perpendicular to the major rostro-caudal ones, but the spatial resolution does not allow to resolve them, and to unambiguously distinguish it from noise. Taking this previous evidence as motivation, the present study used 3D polarized light imaging (3D-PLI) for micrometer resolution analysis of nerve fibers in postmortem specimens of a vervet monkey brain. The analysis of coronal occipital and parietal sections revealed that the sagittal stratum consisted of an external and an internal layer, which are joined and crossed by fibers from the surrounding white matter and the tapetum. Fibers from different parietal and occipital regions entered the sagittal stratum in the dorsal, ventral or middle sector, as solid large bundles or as several small fiber aggregations. These patterns were remarkably similar to published results of tracer experiments in macaques. Taking this correspondence as external validation of 3D-PLI enabled translation to the human brain, where a similarly complex fiber architecture within the sagittal stratum could be exemplified in a human hemisphere in our study. We thus argue in favor of a dedicated fiber microstructure within the sagittal stratum as a correlate of the additional fiber directions typically seen in in-vivo diffusion imaging studies.

Human Achilles tendon mechanical behavior is more strongly related to collagen disorganization than advanced glycation end-products content

Diabetes is associated with impaired tendon homeostasis and subsequent tendon dysfunction, but the mechanisms underlying these associations is unclear. Advanced glycation end-products (AGEs) accumulate with diabetes and have been suggested to alter tendon function. In vivo imaging in humans has suggested collagen disorganization is more frequent in individuals with diabetes, which could also impair tendon mechanical function. The purpose of this study was to examine relationships between tendon tensile mechanics in human Achilles tendon with accumulation of advanced glycation end-products and collagen disorganization. Achilles tendon specimens (n = 16) were collected from individuals undergoing lower extremity amputation or from autopsy. Tendons were tensile tested with simultaneous quantitative polarized light imaging to assess collagen organization, after which AGEs content was assessed using a fluorescence assay. Moderate to strong relationships were observed between measures of collagen organization and tendon tensile mechanics (range of correlation coefficients: 0.570–0.727), whereas no statistically significant relationships were observed between AGEs content and mechanical parameters (range of correlation coefficients: 0.020–0.210). Results suggest that the relationship between AGEs content and tendon tensile mechanics may be masked by multifactorial collagen disorganization at larger length scales (i.e., the fascicle level).

Mechanical properties and microstructural organization of common ulnar collateral ligament grafts: Palmaris longus and gracilis tendons.

Ulnar collateral ligament (UCL) injuries are becoming increasingly common. The palmaris longus (PL) and gracilis (GR) tendons are the most common grafts used in UCL reconstructions. While clinical studies have demonstrated relatively similar outcomes for either graft, there is little quantitative data describing these grafts from a material perspective, specifically the mechanical and microstructural properties of these tissues and how they respond under dynamic loading. The purpose of this descriptive laboratory study was to quantify and compare the mechanical and microstructural properties of PL and GR tendons. A total of 13 PL and 11 GR cadaveric human tendons were obtained. Each specimen was divided into three subregions and subjected to preconditioning, ramp-and-hold stress-relaxation and ramp-to-failure testing. Mechanical parameters were computed for each sample, and a polarized light imaging technique was used to simultaneously evaluate dynamic microstructural properties during testing. The PL had larger toe- and linear-region modulus values than the GR. Within the GR, the distal subregion had stronger collagen alignment than the proximal subregion at the zero, transition and linear portions of the stress-strain curve. The PL and GR, have similar mechanical properties and similar microstructural alignment under load. The PL graft has similar properties throughout its length whereas the GR properties exhibited slight differences in strength of alignment along its length. The PL and GR exhibit larger moduli values and more strongly/uniformly aligned collagenous microstructure when qualitatively compared to data previously published on the native UCL.

Donor age and sex have limited effects on the mechanical and microstructural properties of human connective tissues.

Connective tissues, such as tendons, ligaments, and capsules, play a large role in locomotion and joint stability and are often subjected to traumatic injuries and degeneration. The purpose of this study was to evaluate if the mechanical and microstructural properties of connective tissues correlate with the age and sex of the human donor. Dissected samples were prepared for mechanical testing, consisting of 10 cycles of preconditioning, a stress-relaxation ramp and hold, and a quasi-static ramp to failure. During the testing protocol, the microstructural organization of tissues was analyzed using quantitative polarized light imaging. A linear mixed model was used to assess whether tissue type, donor age, or donor sex were significantly associated with mechanical and microstructural tissue properties. Tissue type had a significant effect on all parameters, while donor age and sex did not. Groupings by tissue type (i.e., tendon vs. ligament vs. capsule) were evident for microstructural data, with tendons having a tighter grouping and ligaments having a larger spread of values. The interaction of tissue type and age yielded a significant effect for linear modulus only (p = 0.007), with the palmaris tendon appearing to have the largest contribution to this effect. There were no significant interaction effects between sex and tissue type or donor age. Donor age appears to affect linear modulus in some, but not all, tissue types. Otherwise, age and sex do not have significant effects on the mechanical and microstructural properties of the range of connective tissues that were analyzed in this study.

Milk fat globule membrane attenuates high fat diet-induced neuropathological changes in obese Ldlr-/-.Leiden mice.

Milk-fat globule membrane (MFGM) is a complex structure secreted by the mammary gland and present in mammalian milk. MFGM contains lipids and glycoproteins as well as gangliosides, which may be involved in myelination processes. Notably, myelination and thereby white matter integrity are often altered in obesity. Furthermore, MFGM interventions showed beneficial effects in obesity by affecting inflammatory processes and the microbiome. In this study, we investigated the impact of a dietary MFGM intervention on fat storage, neuroinflammatory processes and myelination in a rodent model of high fat diet (HFD)-induced obesity. 12-week-old male low density lipoprotein receptor-deficient Leiden mice were exposed to a HFD, a HFD enriched with 3% whey protein lipid concentrate (WPC) high in MFGM components, or a low fat diet. The impact of MFGM supplementation during 24-weeks of HFD-feeding was examined over time by analyzing body weight and fat storage, assessing cognitive tasks and MRI scanning, analyzing myelinization with polarized light imaging and examining neuroinflammation using immunohistochemistry. We found in this study that 24 weeks of HFD-feeding induced excessive fat storage, increased systolic blood pressure, altered white matter integrity, decreased functional connectivity, induced neuroinflammation and impaired spatial memory. Notably, supplementation with 3% WPC high in MFGM components restored HFD-induced neuroinflammation and attenuated the reduction in hippocampal-dependent spatial memory and hippocampal functional connectivity. We showed that supplementation with WPC high in MFGM components beneficially contributed to hippocampal-dependent spatial memory, functional connectivity in the hippocampus and anti-inflammatory processes in HFD-induced obesity in rodents. Current knowledge regarding exact biological mechanisms underlying these effects should be addressed in future studies.