Organization Of Fibers Research Articles

Elucidation of the Origin of Thixotropic-Inducing Properties of Additive Amphiphiles and the Creation of a High-Performance Triamide Amphiphile.

The spontaneous growth of helical fibers of amphiphilic diamide derivatives containing hydrocarbons with asymmetric carbon centers in their constituent hydrocarbons was investigated. 12-Hydroxystearic acid and a gemini-type surfactant obtained by the bimolecular condensation of this compound with hexamethylenediamine both impart thixotropic ability to a solvent. Although this thixotropic behavior is based on the growth of hierarchical crystalline nanofibers in the solvents, the degree of fiber growth itself was not the origin of the thixotropy. In this study, it has adopted the methods of the Langmuir monolayer and Langmuir-Blodgett films as technique to selectively and individually evaluate the behavior of 12-hydroxyl stearyl and/or stearyl chains themselves. The ability to impart thixotropy to the solvent via fiber organization was related to the intermolecular hydrogen bonding between the added amphiphiles. Additionally, homogeneous right-handed helical fibers were formed in the spin-cast films of the diamide derivatives, and a positive Cotton effect was observed in their circular dichroism spectra. It is suggested that fibers that do not form helical arrangements cannot impart sufficient thixotropy to the solvent even when extensive fiber growth is achieved, and the structure-dependent development of chirality is the driving force. In addition, to further the development of highly functional thixotropic agents, a trefoil-like triamide derivative containing three chains was synthesized. By using this molecule, solvent gelation occurred at 78% as an addition to the diamide case, and a supramolecular assembly was formed in the corresponding two-dimensional film.

Biomechanical Characterization of Intracranial Aneurysm Wall: A Multiscale Study

Aneurysm wall biomechanics are not yet an integral part of aneurysm rupture risk evaluation. We aimed to develop a new technique describing the biomechanical properties of aneurysm wall and correlating them to rupture status. Aneurysm wall samples collected during surgery were submitted before and after freezing to tensile tests or as fresh samples to indentation tests. The lateral stiffness or the Young's modulus of the different samples was determined as a function of the mechanical test used. The impact of freezing on biomechanical properties was evaluated. The correlation of clinical and radiologic data with the biomechanical profile of the aneurysm samples was investigated. Two-photon microscopy was used to study collagen fiber organization. Sixteen aneurysm samples (11 unruptured and 5 ruptured) were included. Freezing decreased tissue stiffness. No significant difference was found between ruptured and unruptured aneurysm wall samples regarding demographic characteristics, ethnicity, smoking status, arterial hypertension, site, size and shape of the aneurysm, PHASES score, mechanical profile, or overall Young's modulus. Indentation tests found that the rupture occurred in a restricted area of increased elastic capacity and unruptured areas had increased stiffness. Two-photon microscopy found disruption of the collagen fiber network in rupture zones. The indentation test of fresh aneurysm wall samples described the heterogeneity of biomechanical properties of the tissue and found increased elastic capacity in the rupture zone and increased stiffness in the remainder of the aneurysm. This study could be a basis for further research aimed at building a biomechanical-based model of aneurysm rupture risk.

Structural network topology correlates of microstructural brain dysmaturation in term infants with congenital heart disease.

Neonates with complex congenital heart disease (CHD) demonstrate microstructural brain dysmaturation, but the relationship with structural network topology is unknown. We performed diffusion tensor imaging (DTI) in term neonates with CHD preoperatively (N = 61) and postoperatively (N = 50) compared with healthy term controls (N = 91). We used network topology (graph) analyses incorporating different weighted and unweighted approaches and subject-specific white matter segmentation to investigate structural topology differences, as well as a voxel-based analysis (VBA) to confirm the presence of microstructural dysmaturation. We demonstrate cost-dependent network inefficiencies in neonatal CHD in the pre- and postoperative period compared with controls, related to microstructural differences. Controlling for cost, we show the presence of increased small-worldness (hierarchical fiber organization) in CHD infants preoperatively, that persists in the postoperative period compared with controls, suggesting the early presence of brain reorganization. Taken together, topological microstructural dysmaturation in CHD infants is accompanied by hierarchical fiber organization during a protracted critical period of early brain development. Our methodology also provides a pipeline for quantitation of network topology changes in neonates and infants with microstructural brain dysmaturation at risk for perinatal brain injury.

Mitochondrial dynamics (fission and fusion) and collagen production in a rat model of diabetic wound healing treated by photobiomodulation: comparison of 904 nm laser and 850 nm light-emitting diode (LED)

ObjectiveMitochondrial dysfunction has been associated with the development of diabetes mellitus which is characterized by disorders of collagen production and impaired wound healing. This study analyzed the effects of photobiomodulation (PBM) mediated by laser and light-emitting diode (LED) on the production and organization of collagen fibers in an excisional wound in an animal model of diabetes, and the correlation with inflammation and mitochondrial dynamics. MethodsTwenty Wistar rats were randomized into 4 groups of 5 animals. Groups: (SHAM) a control non-diabetic wounded group with no treatment; (DC) a diabetic wounded group with no treatment; (DLASER) a diabetic wounded group irradiated by 904 nm pulsed laser (40 mW, 9500 Hz, 1 min, 2.4 J); (DLED) a diabetic wounded group irradiated by continuous wave LED 850 nm (48 mW, 22 s, 1.0 J). Diabetes was induced by injection with streptozotocin (70 mg/kg). PBM was carried out daily for 5 days followed by sacrifice and tissue removal. ResultsCollagen fibers in diabetic wounded skin were increased by DLASER but not by DLED. Both groups showed increased blood vessels by atomic force microscopy. Vascular endothelial growth factor (VEGF) was higher and cyclooxygenase (COX2) was lower in the DLED group. Mitochondrial fusion was higher and mitochondrial fusion was lower in DLED compared to DLASER. ConclusionDifferences observed between DLASER and DLED may be due to the pulsed laser and CW LED, and to the higher dose of laser. Regulation of mitochondrial homeostasis may be an important mechanism for PBM effects in diabetes.

A comparative study of the elastic fibre system within the mouse and human cornea

The cornea relies on its organised extracellular matrix for maintaining transparency and biomechanical strength. Studies have identified an elastic fibre system within the human posterior cornea, thought to allow for slight deformations in response to internal pressure fluctuations within the eye. However, the type of elastic fibres that exist within the cornea and their roles remain elusive. The aim of this study was to compare the distribution and organisation of the elastic fibres within the posterior peripheral mouse and human cornea, and elucidate how these fibres integrate with the trabecular meshwork, whilst characterising the distribution of their main likely components (fibrillin-1, elastin and type VI collagen) in different parts of the cornea and adjacent sclera.We identified key differences in the elastic fibre system between the human and mouse cornea. True elastic fibres (containing elastin) were identified within the human posterior peripheral cornea. Elastic fibres appeared to present as an extensive network throughout the mouse corneal stroma, but as fibrillin-rich microfibril bundles rather than true elastic fibres. However, tropoelastin staining indicated the possibility that true elastic fibres had yet to develop in the young mice studied. Differences were also apparent within the anatomy of the trabecular meshwork. The human trabecular meshwork appeared to insert between the corneal stroma and Descemet's membrane, with elastic fibres continuing into the stroma from the trabecular meshwork anterior to Descemet's membrane. Within the mouse cornea, no clear insertion point of the trabecular meshwork was seen, instead the elastic fibres within the trabecular meshwork continued into Descemet's membrane, with the trabecular meshwork joining posterior to Descemet's membrane.

Kartogenin Enhances Collagen Organization and Mechanical Strength of the Repaired Enthesis in a Murine Model of Rotator Cuff Repair

To investigate the use of kartogenin (KGN) in augmenting healing of the repaired enthesis after rotator cuff repair in a murine model. Seventy-two C57BL/6 wild-type mice underwent unilateral detachment and transosseous repair of the supraspinatus tendon augmented with either fibrin sealant (control group; n= 36) or fibrin sealant containing 100μmol/L of KGN (experimental group; n= 36) applied at the repair site. Postoperatively, mice were allowed free cage activity without immobilization. Mice were humanely killed at 2 and 4weeks postoperatively. Repair site integrity was evaluated histologically through fibrocartilage formation and collagen fiber organization and biomechanically through load-to-failure testing of the supraspinatus tendon-bone construct. At 2weeks, no differences were noted in percent area of fibrocartilage, collagen organization, or ultimate strength between groups. At 4weeks, superior collagen fiber organization (based on collagen birefringence [17.3 ± 2.0 vs 7.0 ± 6.5 integrated density/μm2; P<.01]) and higher ultimate failure loads (3.5 ± 0.6N vs 2.3 ± 1.1N; P= .04) were seen in the KGN group. The percent area of fibrocartilage (13.2 ± 8.4% vs 4.4 ± 5.4%; P= .04) was higher in the control group compared with the KGN group. Rotator cuff repair augmentation with KGN improved the collagen fiber organization and biomechanical strength of the tendon-bone interface at 4weeks in a murine model. These findings have implications for improving the structural integrity of the repaired enthesis and potentially reducing the retear rate after rotator cuff repair, which can ultimately lead to improvements in clinical outcomes.

Experimental model of temporomandibular joint arthritis: Evaluation of contralateral joint and masticatory muscles

ObjectiveTo investigate morphological and biochemical changes in an experimental model of temporomandibular joint (TMJ) arthritis, as well as examine contralateral joint involvement and morphometric aspects of masticatory muscles in affected animals. DesignForty Wistar rats were allocated into three groups, as follows: a healthy control group (HG); and a group with arthritis induced in the left TMJ (IG); and a contralateral group with noninflamed right TMJ (CG). Arthritis was induced by intra-articular injection of 50 μL of Complete Freund's Adjuvant (CFA). Morphological analysis was performed using histological sections of TMJs stained with hematoxylin-eosin, picrosirius and toluidine blue. In addition, histomorphometric analyses of the articular cartilage, articular disc and masticatory muscles, were performed. Biochemical analysis of TMJ tissues included dosage of sulfated glycosaminoglycans (GAGs), zymography of the Matrix Metalloproteinases (MMPs) 2 and 9 and ELISA for the cytokines IL-6, TNF-α and IL-1β. The data were analyzed by paired and unpaired t-tests, with a p < 0.05. ResultsIG animals presented morphological and biochemical changes in their TMJ, including synovial membrane hyperplasia; chronic inflammatory infiltrate; collagen fiber disorganization in the disc; reduced proteoglycans in the cartilage; greater disc thickness; lower cartilage thickness; lower feed intake; higher activity of MMP-9 and MMP-2, and higher concentration of IL-6, TNF-α, and IL-1β, as compared to HG. CG animals showed posterior disc thickening; reduced proteoglycans in the cartilage; lower collagen fiber organization of the disc; active MMP-9, and higher concentration of IL-6 and TNF-α compared to HG animals. Articular inflammation reduced the area and diameter of fibers of the temporal (right), medial pterygoid (left), and lateral pterygoid (right and left) muscles as compared to HG animals. ConclusionTMJ arthritis reduced the area and diameter of masticatory muscle fibers and caused morphological and molecular changes in the contralateral joint.

3D skeletal muscle fascicle engineering is improved with TGF-β1 treatment of myogenic cells and their co-culture with myofibroblasts.

BackgroundSkeletal muscle wound healing is dependent on complex interactions between fibroblasts, myofibroblasts, myogenic cells, and cytokines, such as TGF-β1. This study sought to clarify the impact of TGF-β1 signaling on skeletal muscle cells and discern between the individual contributions of fibroblasts and myofibroblasts to myogenesis when in co-culture with myogenic cells. 3D tissue-engineered models were compared to equivalent 2D culture conditions to assess the efficacy of each culture model to predictively recapitulate the in vivo muscle environment.MethodsTGF-β1 treatment and mono-/co-cultures containing human dermal fibroblasts or myofibroblasts and C2C12 mouse myoblasts were assessed in 2D and 3D environments. Three culture systems were compared: cell monolayers grown on 2D dishes and 3D tissues prepared via a self-assembly method or collagen 1-based hydrogel biofabrication. qPCR identified gene expression changes during fibroblast to myofibroblast and myoblast differentiation between culture conditions. Changes to cell phenotype and tissue morphology were characterized via immunostaining for myosin heavy chain, procollagen, and α-smooth muscle actin. Tissue elastic moduli were measured with parallel plate compression and atomic force microscopy systems, and a slack test was employed to quantify differences in tissue architecture and integrity.ResultsTGF-β1 treatment improved myogenesis in 3D mono- and co-cultures containing muscle cells, but not in 2D. The 3D TGF-β1-treated co-culture containing myoblasts and myofibroblasts expressed the highest levels of myogenin and collagen 1, demonstrating a greater capacity to drive myogenesis than fibroblasts or TGF-β1-treatment in monocultures containing only myoblasts. These constructs possessed the greatest tissue stability, integrity, and muscle fiber organization, as demonstrated by their rapid and sustained shortening velocity during slack tests, and the highest Young’s modulus of 6.55 kPA, approximate half the stiffness of in situ muscle. Both self-assembled and hydrogel-based tissues yielded the most multinucleated, elongated, and aligned muscle fiber histology. In contrast, the equivalent 2D co-culture model treated with TGF-β1 completely lacked myotube formation through suppression of myogenin gene expression.DiscussionThese results show skeletal muscle regeneration can be promoted by treating myogenic cells with TGF-β1, and myofibroblasts are superior enhancers of myogenesis than fibroblasts. Critically, both TGF-β1 treatment and co-culturing skeletal muscle cells with myofibroblasts can serve as myogenesis accelerators across multiple tissue engineering platforms. Equivalent 2D culture systems cannot replicate these affects, however, highlighting a need to continually improve in vitro models for skeletal muscle development, discovery of therapeutics for muscle regeneration, and research and development of in vitro meat products.

Abstract 2114: 3D co-culture of pancreatic tumor sphroids and stellate cells using minipillar array as a microtumor model of pancreatic ductal adenocarcinoma

Abstract Purpose: Tumor microenvironmental (TME) factors play important roles in cancer progression and chemoresistance by inducing epithelial-mesenchymal transition (EMT) and related mechanisms. Pancreatic stellate cell (PSC) is a highly-enriched cell type in TME of pancreatic ductal adenocarcinoma (PDAC). Three dimensional (3D) tumor spheroids (TS) are considered as an in vivo-like tumor model demonstrating its clinical relevancy. We previously developed a minipillar array histochip for 3D culture and immunohistological analysis of TS in an increased throughput manner. Here, we present a minipillar-based co-culture of TS and PSC as a novel in vitro microtumor model of pancreatic ductal adenocarcinoma. Methods: Minipillar chips were obtained from MBD Co. (Cellvitro™ 55Cryo). 3D TS of Panc-1, human PDAC cells and PSCs (Sciencell) were cultured on minipillars and at the bottom of 96 well plates, respectively, as embedded in type I collagen. Changes in expression of extracellular matrix (ECM) and EMT-related factors in TS were assessed by confocal microscopy following cyrosection preparation. Results: Co-culture condition was optimized for the growth of TS, activation of PSC, and their interaction. Expression of EMT markers such as TGF-β1 and vimentin increased in co-cultured TS than mono-cultured TS. ECM remodeling as indicated by increased matrix deposition (collagen I, fibronectin) and fiber organization was observed in TS co-cultured with PSC compared to mono-cultured TS. Upon drug exposure (gemcitabine and paclitaxel), survival advantage was prominent in TS co-cultured with activated PSC. Conclusions: We established 3D model of PDAC by co-culturing TS and PSC using minipillar histochips. EMT-related changes including ECM remodeling and drug resistance were recapitulated in the model. Overall our results indicate usefulness of the present model not only in studying the role of TME factors and their interactions towards tumor progression but also in the evaluation of therapeutic efficacy of drug candidates and combinations. Citation Format: Hyun Ju Hwang, Min-Suk Oh, Seul-Ki Kim, Hyo-Jeong Kuh. 3D co-culture of pancreatic tumor sphroids and stellate cells using minipillar array as a microtumor model of pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2114.

Abstract 3598: Classification of the collagen ECM in normal human tissue as a biomarker for future breast cancer incidence

Abstract Because we do not know who will develop breast cancer, or who will have a relapse of cancer following surgery, there is an urgent need for the development of biomarkers that can be detected specifically in normal, nondiseased tissue. For 30 years there has been a search for the genetic markers that will predict risk of developing breast cancer, but even while family history is a known risk factor, only 10% of breast cancer risk can be specifically linked to a genetic mechanism (BRCA-1, BRCA-2). Furthermore, individuals who are at high risk due to genetics or family history do not always develop breast cancer. Breast density is another indicator of future incidence, as high-density tissue is correlated with a 4-6 fold increased risk of developing cancer. We hypothesized that a population analysis of the properties of the individual collagen fibers that comprise the extracellular matrix (ECM) in nondiseased breast tissue is a biomarker for the onset of disease. We used multiphoton second-harmonic generation (SHG) imaging to generate high-contrast images of the collagen ECM that can be gathered from common histologic slides that require no additional staining. Our goal was to determine the range of heterogeneity in the properties of the collagen matrix in normal samples. We imaged samples from 141 normal patient donors to comprise 4 different cohorts that define this ground truth state of the matrix. The cohorts were from donors who had never been diagnosed with breast cancer; who had previously been diagnosed with breast cancer but now were disease free; who had never been diagnosed with breast cancer at the time of tissue donation but later went on to develop breast cancer; and those who had donated tissue while in remission from an initial breast cancer diagnosis but later went on to have a recurrence of breast cancer. The SHG images of each patient sample were then analyzed using the curvelet transform-based ctFIRE and CurveAlign software platforms to generate data (fiber length, width, straightness, density, angle with respect to a boundary of epithelial ducts/lobules, and the relative alignment of fibers to each other) on each individual collagen fiber. Advanced statistical measurements and principal component analysis were used to classify the nature of each of the cohorts. Based on our measurements of the structure and organization of collagen fibers, we found that while measurements within a cohort were consistent, there were unique attributes of the collagen matrix that defined each cohort. Furthermore, we performed regression analysis of our measurements against the standard clinical features (age, race, BMI, etc.). Because our various cohorts delineate the nondiseased, involuted, precancer, and prerecurrence normal matrix, respectively, we feel that these data serve as a highly useful, novel classifier, one that describes the clinical impact of the timbre of the matrix in nondiseased women. Citation Format: Ryan J. Gigstad, Tianjie Wang, Yifei Liu, Menggang Yu, Yuming Liu, Adib Keikhosravi, Kevin W. Eliceiri, Patricia J. Keely, Matthew W. Conklin. Classification of the collagen ECM in normal human tissue as a biomarker for future breast cancer incidence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3598.

Characterization of local muscle fiber anisotropy using shear wave elastography in patients with chronic myofascial pain

Myofascial trigger points (MTrPs) are common in soft tissue musculoskeletal pain conditions. It is believed that MTrPs are local contractures within the extrafusal fibers of skeletal muscles. Further characterizing muscle pathophysiology is desirable to design effective treatments. This study was designed to measure muscle fiber architecture in the neighbourhood of MTrPs using ultrasound Shear Wave Elastography (SWE), a novel approach for quantitatively assessing muscle fiber. We hypothesized that muscle containing active (symptomatic) MTrPs has disorganized fiber architecture. Twenty-four participants (14 women, ages 20–60 years) met criteria for chronic myofascial pain affecting the neck region. All underwent a clinical history and physical examination of the neck and upper trapezius to identify MTrPs and pain self-report. A Supersonic Aixplorer ultrasound imaging system with an L10-4 ultrasound transducer was used to image the upper trapezius muscle. A custom transducer holder ( Fig. 1 ) enabled imaging through a 20 mm diameter window placed over the palpable MTrP or normal tissue, and rotation of the imaging plane to precise angles from 0° (along fibers) to 90° (perpendicular to fibers). Peak shear wave velocity (SWV) is observed longitudinally along muscle fibers (0°), and is minimum across fibers (90°). Muscle fiber anisotropy (a measure of fiber alignment and organization) was quantified as the asymmetry (degrees) in the shear-wave velocity profile around the peak SWV. Muscles with active (symptomatic) MTrPs ( n = 24) exhibited greater anisotropy (14.25 ± 11.01°), compared to normal, asymptomatic muscle tissue ( n = 12), which is more isotropic (7.16 ± 6.70°) ( P < 0.05) ( Fig. 2 ). A positive correlation was found between the maximum anisotropy and average pain (Pearson's r = 0.54). Muscle containing active MTrPs is anisotropic compared to normal muscle tissue, suggesting muscle contractures are in regions of anisotropy. The degree of anisotropy correlates with pain levels.

Topical retinol attenuates stress-induced ageing signs in human skin ex vivo, throughEGFR activation viaEGF, but notERK andAP-1 activation.

Stress-induced oxidative damage and the inflammatory response lead to degradation of collagen and elastic fibres and wrinkle formation. Topical retinol (or vitamin A) can be a strategy to attenuate the effects of stress in skin as it promotes collagen and elastic fibre production and reduces protease synthesis. This study investigated the effect of topical retinol in stressed human skin using in vitro and ex vivo models. Human skin explants were treated with high levels of epinephrine (as observed in stressed patients) and topically with retinol for 13days. Human dermal fibroblasts were treated with conditioned medium of ex vivo retinol-treated and non-stressed (without epinephrine) human skin for 24hours. In ex vivo human skin, retinol reversed the epinephrine-induced reduction in epidermal proliferation and differentiation, normalizing epidermal thickness. Retinol also inhibited the epinephrine-induced reduction in elastic fibre deposition and organization, restoring dermal thickness. In addition, retinol reversed the epinephrine-induced increase in c-JUN protein expression, but it did not alter extracellular signal-regulated kinase 1/2 (ERK) phosphorylation in ex vivo human skin. Conditioned medium of ex vivo retinol-treated and non-stressed human skin presented an increased protein expression of epidermal growth factor (EGF). In human dermal fibroblasts, conditioned medium of ex vivo retinol-treated and non-stressed human skin increased protein and gene expression of fibrillin-1 and protein expression of EGF receptor (EGFR). In conclusion, topical retinol attenuates stress-induced skin ageing signs in human skin ex vivo, probably through EGFR activation via EGF, but not by the stress-activated ERK 1/2 and c-JUN pathways.

3D organizational mapping of collagen fibers elucidates matrix remodeling in a hormone-sensitive 3D breast tissue model

Hormones play an important role in normal and diseased breast tissue development. However, they can also disrupt cell-matrix interactions and their role in extracellular matrix reorganization during epithelial morphogenesis remains poorly understood, partly due to a lack of sensitive approaches for matrix characterization. Here, we assess the hormonal regulation of matrix reorganization in a three-dimensional (3D) breast tissue culture model using a novel metric, i.e., 3D directional variance, to characterize the 3D organization of collagen fibers visualized via high-resolution, second harmonic generation imaging. This metric enables resolving and quantifying patterns of spatial organization throughout the matrix surrounding epithelial structures treated with 17β-estradiol (E2) alone, and E2 in combination with either promegestone, a progestogen, or prolactin. Addition of promegestone results in the most disorganized fibers, while the E2 alone treatment leads to the most organized ones. Location-dependent organization mapping indicates that only the prolactin treatment leads to significant heterogeneities in the regional organization of collagen fibers, with higher levels of alignment observed at the end of the elongated epithelial structures. The observed collagen organization patterns for all groups persist for tens of micrometers. In addition, a comparison between 3D directional variance and typical 2D analysis approaches reveals an improved sensitivity of the 3D metric to identify organizational heterogeneities and differences among treatment groups. These results demonstrate that 3D directional variance is sensitive to subtle changes in the extracellular micro-environment and has the potential to elucidate reciprocal cell-matrix interactions in the context of numerous applications involving the study of normal and diseased tissue morphogenesis.

Pleistocene sea‐floor fibrous crusts and spherulites in the Danakil Depression (Afar, Ethiopia)

AbstractPleistocene fibrous aragonite fabrics, including crusts and spherules, occur in the Danakil Depression (Afar, Ethiopia) following the deposition of two distinctive Middle and Late Pleistocene coralgal reef units and pre‐dating the precipitation of evaporites. Crusts on top of the oldest reef unit (Marine Isotope Stage 7) cover and fill cavities within a red algal framework. The younger aragonite crusts directly cover coralgal bioherms (Marine Isotope Stage 5) and associated deposits. Their stratigraphic position between marine and evaporitic deposits, and their association to euryhaline molluscs, suggest that the crusts and spherules formed in restricted semi‐enclosed conditions. The availability of hard substrate controls crust formation with crusts more often found on steep palaeo‐slopes, from sea level up to at least 80 m depth, while spherules mainly occur associated with mobile substrate. Crusts reach up to 30 cm in thickness and can be microdigitate, columnar (branching and non‐branching) or non‐columnar, with laminated and non‐laminated fabrics. Two different lamination types are found within the crystalline fabrics: (i) isopachous lamination; and (ii) irregular lamination. These two types of lamination can be distinguished by the organization of the aragonite fibres, as well as the lateral continuity of the laminae. Scanning electron microscopy with energy dispersive X‐ray spectroscopy analyses on well‐preserved samples revealed the presence of Mg‐silicate laminae intercalated with fibrous aragonite, as well as Mg‐silicate aggregates closely associated with the fibrous aragonite crusts and spherules. The variety of observed fabrics results from a continuum of abiotic and microbial processes and, thus, reflects the tight interaction between microbially mediated and abiotic mineralization mechanisms. These are the youngest known isopachously laminated, digitate and columnar branching fibrous crusts associated with a transition from marine to evaporitic conditions. Understanding the context of formation of these deposits in Afar can help to better interpret the depositional environment of the widespread Precambrian sea‐floor precipitates.

Daily oral supplementation with collagen peptides combined with vitamins and other bioactive compounds improves skin elasticity and has a beneficial effect on joint and general wellbeing

Aging is a multifactorial and natural process that causes physiological changes in organs, tissues and cells over time. In the skin and cartilage, aging leads to a decrease in the synthesis and changes in the arrangement of proteoglycans and collagen, in addition to the loss of glycosaminoglycans, which are responsible for the integrity and health of these tissues. We hypothesized that daily oral supplementation with a liquid nutraceutical containing hydrolyzed fish collagen, vitamins, antioxidants and other active ingredients could improve skin texture and elasticity, and in addition have a protective effect on joint health. A double-blind, randomized, placebo-controlled clinical trial was conducted on 120 subjects who consumed either the test product or placebo on a daily basis for 90 days. Subjects consuming the test product had an overall significant increase in skin elasticity (+40%; P < .0001) when compared to placebo. Histological analysis of skin biopsies revealed positive changes in the skin architecture, with a reduction in solar elastosis and improvement in collagen fiber organization in the test product group. As reported in the self-perception questionnaires, these results were confirmed by the subjects' own perceptions in that participants agreed their skin was more hydrated and more elastic. In addition, the consumption of the test product reduced joint pain by −43% and improved joint mobility by +39%. Oral supplementation with collagen bioactive peptides combined with chondroitin sulphate, glucosamine, L-carnitine, vitamins, and minerals significantly improved the clinical parameters related to skin aging and joint health, and therefore, might be an effective solution to slow down the hallmarks of aging.

Keratinocyte Motility Is Affected by UVA Radiation-A Comparison between Normal and Dysplastic Cells.

UVA radiation induces multiple and complex changes in the skin, affecting epidermal cell behavior. This study reports the effects of UVA exposure on normal (HaCaT) and dysplastic (DOK) keratinocytes. The adherence, spreading and proliferation were investigated by time-lapse measurement of cell layer impedance on different matrix proteins. Prior to UVA exposure, the time required for adherence and spreading did not differ significantly for HaCaT and DOK cells, while spreading areas were larger for HaCaT cells. Under UVA exposure, HaCaT and DOK cells behavior differed in terms of movement and proliferation. The cells’ ability to cover the denuded surface and individual cell trajectories were recorded by time-lapse videomicroscopy, during wound healing experiments. Dysplastic keratinocytes showed more sensitivity to UVA, exhibiting transient deficiencies in directionality of movement and a delay in re-coating the denuded area. The actin cytoskeleton displayed a cortical organization immediately after irradiation, in both cell lines, similar to mock-irradiated cells. Post-irradiation, DOK cells displayed a better organization of stress fibers, persistent filopodia, and new, stronger focal contacts. In conclusion, after UVA exposure HaCaT and DOK cells showed a different behavior in terms of adherence, spreading, motility, proliferation, and actin cytoskeleton dynamics, with the dyplastic keratinocytes being more sensitive.

Quantitative analysis of the effect of environmental-scanning electron microscopy on collagenous tissues

Environmental-scanning electron microscopy (ESEM) is routinely applied to various biological samples due to its ability to maintain a wet environment while imaging; moreover, the technique obviates the need for sample coating. However, there is limited research carried out on electron-beam (e-beam) induced tissue damage resulting from using the ESEM. In this paper, we use quantitative second-harmonic generation (SHG) microscopy to examine the effects of e-beam exposure from the ESEM on collagenous tissue samples prepared as either fixed, frozen, wet or dehydrated. Quantitative SHG analysis of tissues, before and after ESEM e-beam exposure in low-vacuum mode, reveals evidence of cross-linking of collagen fibers, however there are no structural differences observed in fixed tissue. Meanwhile wet-mode ESEM appears to radically alter the structure from a regular fibrous arrangement to a more random fiber orientation. We also confirm that ESEM images of collagenous tissues show higher spatial resolution compared to SHG microscopy, but the relative tradeoff with collagen specificity reduces its effectiveness in quantifying collagen fiber organization. Our work provides insight on both the limitations of the ESEM for tissue imaging, and the potential opportunity to use as a complementary technique when imaging fine features in the non-collagenous regions of tissue samples.

Cancer Stem Cell Migration in Three-Dimensional Aligned Collagen Matrices.

Cell migration is strongly influenced by the organization of the surrounding 3-D extracellular matrix. In particular, within fibrous solid tumors, carcinoma cell invasion may be directed by patterns of aligned collagen in the extra-epithelial space. Thus, studying the interactions of heterogeneous populations of cancer cells that include the stem/progenitor-like cancer stem cell subpopulation and aligned collagen networks is critical to our understanding of carcinoma dissemination. Here, we describe a robust method to generate aligned collagen matrices in vitro that mimic in vivo fiber organization. Subsequently, a protocol is presented for seeding aligned matrices with distinct carcinoma cell subpopulations and performing live cell imaging and quantitative analysis of cell migration. Together, the engineered constructs and the imaging techniques laid out here provide a platform to study cancer stem cell migration in 3-D anisotropic collagen with real-time visualization of cellular interactions with the fibrous matrix. © 2018 by John Wiley & Sons, Inc.

Supramolecular poly(acrylic acid)/F127 hydrogel with hydration-controlled nitric oxide release for enhancing wound healing

Topical nitric oxide (NO) delivery has been shown to accelerate wound healing. However, delivering NO to wounds at appropriate rates and doses requires new biomaterial-based strategies. Here, we describe the development of supramolecular interpolymer complex hydrogels comprising PEO-PPO-PEO (F127) micelles embedded in a poly(acrylic acid) (PAA) matrix, with S-nitrosoglutathione (GSNO) molecules dissolved in the hydrophilic domain. We show that PAA:F127/GSNO hydrogels start releasing NO upon hydration at rates controlled by their rates of water absorption. SAXS measurements indicate that the supramolecular structure of the hydrogels retains long-range order domains of F127 micelles. The PAA/F1227 hydrogels displayed dense morphologies and reduced rates of hydration. The NO release rates remain constant over the first 200 min, are directly correlated with the hydration rates of the PAA:F127/GSNO hydrogels, and can be modulated in the range of 40 nmol/g h to 1.5 μmol/g h by changing the PAA:F127 mass ratio. Long-term NO-release profiles over 5 days are governed by the first-order exponential decay of GSNO, with half-lives in the range of 0.5–3.4 days. A preliminary in vivo study on full-thickness excisional wounds in mice showed that topical NO release from the PAA:F127/GSNO hydrogels is triggered by exudate absorption and leads to increased angiogenesis and collagen fiber organization, as well as TGF-β, IGF-1, SDF-1, and IL-10 gene expressions in the cicatricial tissue. In summary, these results suggest that hydration-controlled NO release from topical PAA:F127/GSNO hydrogels is a potential strategy for enhancing wound healing. Statement of SignificanceThe topical delivery of nitric oxide (NO) to wounds may provide significant beneficial results and represent a promising strategy to treat chronic wounds. However, wound dressings capable of releasing NO after application and allowing the modulation of NO release rates, demand new platforms. Here, we describe a novel strategy to overcome these challenges, based on the use of supramolecular poly(acrylic acid) (PAA):F127 hydrogels charged with the NO donor S-nitrosoglutathione (GSNO) from whereby the NO release can be triggered by exudate absorption and delivered to the wound at rates controlled by the PAA:F127 mass ratio. Preliminary in vivo results offer a proof of concept for this strategy by demonstrating increased angiogenesis; collagen fibers organization; and TGF-β, IGF-1, SDF-1, and IL-10 gene expressions in the cicatricial tissue after topical treatment with a PAA:F127/GSNO hydrogel.

Cortical fibers orientation mapping using in-vivo whole brain 7 T diffusion MRI

Diffusion MRI of the cortical gray matter is challenging because the micro-environment probed by water molecules is much more complex than within the white matter. High spatial and angular resolutions are therefore necessary to uncover anisotropic diffusion patterns and laminar structures, which provide complementary (e.g. to anatomical and functional MRI) microstructural information about the cortex architectonic. Several ex-vivo and in-vivo MRI studies have recently addressed this question, however predominantly with an emphasis on specific cortical areas. There is currently no whole brain in-vivo data leveraging multi-shell diffusion MRI acquisition at high spatial resolution, and depth dependent analysis, to characterize the complex organization of cortical fibers. Here, we present unique in-vivo human 7T diffusion MRI data, and a dedicated cortical depth dependent analysis pipeline. We leverage the high spatial (1.05 mm isotropic) and angular (198 diffusion gradient directions) resolution of this whole brain dataset to improve cortical fiber orientations mapping, and study neurites (axons and/or dendrites) trajectories across cortical depths. Tangential fibers in superficial cortical depths and crossing fiber configurations in deep cortical depths are identified. Fibers gradually inserting into the gyral walls are visualized, which contributes to mitigating the gyral bias effect. Quantitative radiality maps and histograms in individual subjects and cortex-based aligned datasets further support our results.