Organization Of Fibers Research Articles

Investigations of Processing-Induced Structural Changes in Horse Type-I Collagen at Sub and Supramolecular Levels.

The aim of this work is to evaluate the effects of different extraction and material processing protocols on the collagen structure and hierarchical organization of equine tendons. Wide and Small Angle X-ray Scattering investigations on raw powders and thin films revealed that not only the extraction and purification treatments, but also the processing conditions may affect the extent of the protein crystalline domain and induce a nanoscale “shield effect.” This is due to the supramolecular fiber organization, which protects the atomic scale structure from the modifications that occur during fabrication protocols. Moreover, X-ray analyses and Fourier Transform Infrared spectroscopy performed on the biomaterial sheds light on the relationship between processing conditions, triple helical content and the organization in atomic and nanoscale domains. It was found that the mechanical homogenization of the slurry in acidic solution is a treatment that ensures a high content of super-organization of collagen into triple helices and a lower crystalline domain in the material. Finally, mechanical tensile tests were carried out, proving that the acidic solution is the condition which most enhances both mechanical stiffness and supramolecular fiber organization of the films.

Cardiac Fibroblasts and the Extracellular Matrix in Regenerative and Nonregenerative Hearts

During the postnatal period in mammals, the heart undergoes significant remodeling and cardiac cells progressively lose their embryonic characteristics. At the same time, notable changes in the extracellular matrix (ECM) composition occur with a reduction in the components considered facilitators of cellular proliferation, including fibronectin and periostin, and an increase in collagen fiber organization. Not much is known about the postnatal cardiac fibroblast which is responsible for producing the majority of the ECM, but during the days after birth, mammalian hearts can regenerate after injury with only a transient scar formation. This phenomenon has also been described in adult urodeles and teleosts, but relatively little is known about their cardiac fibroblasts or ECM composition. Here, we review the pre-existing knowledge about cardiac fibroblasts and the ECM during the postnatal period in mammals as well as in regenerative environments.

The crural interosseous membrane re-visited: a histological and microscopic study.

The aim of this study was to characterize the microscopic structure and sensory nerve endings of the crural interosseous membrane (IM). 13 IMs from 7 cadavers were used to analyze the organization of the collagen fibers, IM’s thickness, distribution of elastic fibers and nerve elements. The IM is mainly a two-layer collagen fascicle structure with the collagen fibers of adjacent layers orientated along different directions, forming angles of 30.5 +/- 1.7° at proximal and 26.6 +/- 2.1° at distal part (P>0.05). The percentage of elastic fibers between the two layers and inside the collagen fascicle layer is 10.1 +/- 0.5% and 2.2 +/- 0.1% (P<0.001). The IM’s thickness at proximal, middle, and distal parts is 268.5 +/- 18.6μm; 293.2 +/- 12.5μm; 365.3 +/- 19.3 μm, respectively (Proximal vs Distal: P<0.001; Middle vs Distal: P<0.05). Nerve elements were present and located both inside and on the surface of the IM, whereas the mechanoreceptors are mainly located on the surface of the IM. Free nerve endings (33.3 +/- 5.0/cm2) and Ruffini corpuscles (3.4 +/- 0.6/cm2) were the predominant sensory elements, while Pacinian corpuscles (1.3 +/- 0.7/cm2) were rarely found. The type of mechanoreceptors found suggests that the IM may play a role in proprioception.

Fibrous bone tissue engineering scaffolds prepared by wet spinning of PLGA.

Having a self-healing capacity, bone is very well known to regenerate itself without leaving a scar. However, critical size defects due to trauma, tumor, disease, or infection involve bone graft surgeries in which complication rate is relatively at high levels. Bone tissue engineering appears as an alternative for grafting. Fibrous scaffolds are useful in tissue engineering applications since they have a high surface-to-volume ratio, and adjustable, highly interconnected porosity to enhance cell adhesion, survival, migration, and proliferation. They can be produced in a wide variety of fiber sizes and organizations. Wet spinning is a convenient way to produce fibrous scaffolds with consistent fiber size and good mechanical properties. In this study, a fibrous bone tissue engineering scaffold was produced using poly(lactic-co-glycolic acid) (PLGA). Different concentrations (20%, 25%, and 30%) of PLGA (PLA:PGA 75:25) (Mw = 66,000-107,000) were wet spun using coagulation baths composed of different ratios (75:25, 60:40, 50:50) of isopropanol and distilled water. Scanning electron microscopy (SEM) and in vitro degradation studies were performed to characterize the fibrous PLGA scaffolds. Mesenchymal stem cells were isolated from rat bone marrow, characterized by flow cytometry and seeded onto scaffolds to determine the most appropriate fibrous structure for cell proliferation. According to the results of SEM, degradation studies and cell proliferation assay, 20% PLGA wet spun in 60:40 coagulation bath was selected as the most successful condition for the preparation of wet-spun scaffolds. Wet spinning of different concentrations of PLGA (20%, 25%, 30%) dissolved in dichloromethane using different isopropanol:distilled water ratios of coagulation baths (75:25, 60:40, 50:50) were shown in this study.

Polypropylene meshes coated with chitosan/polyethylene glycol for the reconstruction of the abdominal wall: an experimental study in rats

ABSTRACT The aim of this study was to characterize the tissue reactions triggered by the polypropylene mesh coated with chitosan and polyethylene glycol film, and if it’s able to prevent the formation of peritoneal adhesions. Defects in the abdominal wall of rats were induced and polypropylene meshes coated with chitosan/polyethylene glycol (CPEG group, n= 12) and uncoated (PP control group, n= 12) were implanted. On the fourth and forty-fifth postoperative day the formation of adhesion and the tissue reaction to the biomaterial was evaluated through histological and histochemical analysis. The area (P= 0.01) and severity (P= 0.002) of the adhesion was significatively less in the CPEG group. On the fourth day the foreign body reaction was less intense in CPEG group (P= 0.018) and the production of collagen fibers was more intense in this group (P= 0.041). The tissue reactions caused by the biomaterials were similar on the 45th day, with the exception of the high organization of collagen fibers in the CPEG group. The CPEG meshes did not fully prevent the formation of adhesions, but minimized the severity of the process. The foreign body reaction promoted by polypropylene meshes coated with CPEG is less intense than that triggered by uncoated polypropylene meshes.

Ventral Precentral Fiber Intersection Area: A Central Hub in the Connectivity of Perisylvian Associative Tracts

The ventral part of the precentral gyrus is considered one of the most eloquent areas. However, little is known about the white matter organization underlying this functional hub. To analyze the subcortical anatomy underlying the ventral part of the precentral gyrus, ie, the ventral precentral fiber intersection area (VPFIA). Eight human hemispheres from cadavers were dissected, and 8 healthy hemispheres were studied with diffusion tensor imaging tractography. The tracts that terminate at the ventral part of the precentral gyrus were isolated. In addition, 6 surgical cases with left side gliomas close to the VPFIA were operated awake with intraoperative electrical stimulation mapping. The connections within the VPFIA are anatomically organized along an anteroposterior axis: the pyramidal pathway terminates at the anterior bank of the precentral gyrus, the intermediate part is occupied by the long segment of the arcuate fasciculus, and the posterior bank is occupied by the anterior segment of the arcuate fasciculus. Stimulation of the VPFIA elicited speech arrest in all cases. The present study shows strong arguments to sustain that the fiber organization of the VPFIA is different from the classical descriptions, bringing new light for understanding the functional role of this area in language. The VPFIA is a critical neural epicenter within the perisylvian network that may represent the final common network for speech production, as it is strategically located between the termination of the dorsal stream and the motor output cortex that directly control speech muscles.

YAP1 is involved in replenishment of granule cell precursors following injury to the neonatal cerebellum

The cerebellum undergoes major rapid growth during the third trimester and early neonatal stage in humans, making it vulnerable to injuries in pre-term babies. Experiments in mice have revealed a remarkable ability of the neonatal cerebellum to recover from injuries around birth. In particular, recovery following irradiation-induced ablation of granule cell precursors (GCPs) involves adaptive reprogramming of Nestin-expressing glial progenitors (NEPs). Sonic hedgehog signaling is required for the initial step in NEP reprogramming; however, the full spectrum of developmental signaling pathways that promote NEP-driven regeneration is not known. Since the growth regulatory Hippo pathway has been implicated in the repair of several tissue types, we tested whether Hippo signaling is involved in regeneration of the cerebellum. Using mouse models, we found that the Hippo pathway transcriptional co-activator YAP1 (Yes-associated protein 1) but not TAZ (transcriptional coactivator with PDZ binding motif, or WWTR1) is required in NEPs for full recovery of cerebellar growth following irradiation one day after birth. Although Yap1 plays only a minor role during normal development in differentiation of NEPs or GCPs, the size of the cerebellum, and in particular the internal granule cell layer produced by GCPs, is significantly reduced in Yap1 mutants after irradiation, and the organization of Purkinje cells and Bergmann glial fibers is disrupted. The initial proliferative response of Yap1 mutant NEPs to irradiation is normal and the cells migrate to the GCP niche, but subsequently there is increased cell death of GCPs and altered migration of granule cells, possibly due to defects in Bergmann glia. Moreover, loss of Taz along with Yap1 in NEPs does not abrogate regeneration or alter development of the cerebellum. Our study provides new insights into the molecular signaling underlying postnatal cerebellar development and regeneration.

Combined multiphoton imaging and biaxial tissue extension for quantitative analysis of geometric fiber organization in human reticular dermis

The geometric organization of collagen fibers in human reticular dermis and its relationship to that of elastic fibers remain unclear. The tight packing and complex intertwining of dermal collagen fibers hinder accurate analysis of fiber orientation. We hypothesized that combined multiphoton microscopy and biaxial extension could overcome this issue. Continuous observation of fresh dermal sheets under biaxial extension revealed that the geometry of the elastic fiber network is maintained during expansion. Full-thickness human thigh skin samples were biaxially extended and cleared to visualize the entire reticular dermis. Throughout the dermis, collagen fibers straightened with increased inter-fiber spaces, making them more clearly identifiable after extension. The distribution of collagen fibers was evaluated with compilation of local orientation data. Two or three modes were confirmed in all superficial reticular layer samples. A high degree of local similarities in the direction of collagen and elastic fibers was observed. More than 80% of fibers had directional differences of ≤15°, regardless of layer. Understanding the geometric organization of fibers in the reticular dermis improves the understanding of mechanisms underlying the pliability of human skin. Combined multiphoton imaging and biaxial extension provides a research tool for studying the fibrous microarchitecture of the skin.

IFT88 controls NuMA enrichment at k-fibers minus-ends to facilitate their re-anchoring into mitotic spindles

To build and maintain mitotic spindle architecture, molecular motors exert spatially regulated forces on microtubules (MT) minus-ends. This spatial regulation is required to allow proper chromosomes alignment through the organization of kinetochore fibers (k-fibers). NuMA was recently shown to target dynactin to MT minus-ends and thus to spatially regulate dynein activity. However, given that k-fibers are embedded in the spindle, our understanding of the machinery involved in the targeting of proteins to their minus-ends remains limited. Intraflagellar transport (IFT) proteins were primarily studied for their ciliary roles but they also emerged as key regulators of cell division. Taking advantage of MT laser ablation, we show here that IFT88 concentrates at k-fibers minus-ends and is required for their re-anchoring into spindles by controlling NuMA accumulation. Indeed, IFT88 interacts with NuMA and is required for its enrichment at newly generated k-fibers minus-ends. Combining nocodazole washout experiments and IFT88 depletion, we further show that IFT88 is required for the reorganization of k-fibers into spindles and thus for efficient chromosomes alignment in mitosis. Overall, we propose that IFT88 could serve as a mitotic MT minus-end adaptor to concentrate NuMA at minus-ends thus facilitating k-fibers incorporation into the main spindle.

Morphological and histological characterization of an ectopically mineralized structure in a gilthead sea bream Sparus aurata with opercular deformation.

In this study, we describe an abnormal ectopically mineralized structure (EMS) that was found inside the skull of a juvenile Sparus aurata that also showed a bilateral opercular deformation. The overall phenotype and tissue composition were studied using micro-CT scanning and histological analyses. The ectopic structure occupies a large volume of the brain cavity, partially extruding into the gill cavity. It shows a dense mineralization and an extracellular matrix-rich phenotype, with variation in both the morphology and size of the cell lacunae, combined with an irregular fibre organization inside the matrix. This study is the first to report such an EMS in a juvenile teleost fish, where the tissue does not resemble any other connective tissue type described in bony fish so far. The tissue phenotype seems to rule out that the EMS corresponds to a tumorous cartilage. Yet, it is rather reminiscent of a highly mineralized structure found in cartilaginous fish, where it is suggested to be associated with damage repair.

Simultaneous tractography and elastography imaging of the zone-specific structural and mechanical responses in articular cartilage under compressive loading.

We quantified the precise zonal cartilage structural and mechanical responses to unconfined compressive loading by using simultaneous PSOCT based optical tractography and elastography imaging. Twelve bovine knee articular cartilage samples from six animals were imaged under bulk compression from 4% to 20%. The results revealed strong evidence that the conventional radial zone could be divided into two sub-zones with distinct mechanical properties. The "upper" part of the radial zone played a critical role in "absorbing" the mechanical compression. The study also showed that the zonal fiber organization greatly affected the cartilage structural and mechanical responses. A strong correlation was observed between the optical birefringence and logarithm of the Young's modulus. These new results provide useful information for improving mechanical modeling of articular cartilage and developing better cartilage-mimetic biomaterials.

Niobo-phosphate bioactive glass films produced by pulsed laser deposition on titanium surfaces for improved cell adhesion

Abstract Implant bioactivity is related to its ability to stimulate the adjacent bone tissue to create strong and direct chemical bonds with its surface, reducing implant failure and total rehabilitation time. Surface treatments are a way to obtain such surface properties. In this work, niobo-phosphate bioactive glass thin films were produced by pulsed laser deposition with two different laser pulse energies (50 and 100 mJ). The chemical composition, crystallinity, surface morphology, and wettability of the films was investigated. In addition, proliferation and cell morphology tests were also performed to evaluate the cell behaviour and cell-surface interaction with these coatings in comparison with pure titanium surface. The deposited films presented an amorphous structure. Target and films showed the same chemical composition. Coated surface showed higher wettability, with lower contact angles, suggesting a hydrophilicity of the samples. In vitro assays presented an increase in the cell number after the first 48 h at the surface coated with higher energy (100 mJ) and a better cell morphology, when focal contact points, cell size and organization of the stress fibers were taking into account. As a conclusion, same chemical composition was observed at target and films and also could be suggested that the coated surfaces, especially the ones with higher energy presented better surface properties and cellular behaviour than the control samples.

Acoustic modification of collagen hydrogels facilitates cellular remodeling.

Developing tunable biomaterials that have the capacity to recreate the physical and biochemical characteristics of native extracellular matrices (ECMs) with spatial fidelity is important for a variety of biomedical, biological, and clinical applications. Several factors have made the ECM protein, collagen I, an attractive biomaterial, including its ease of isolation, low antigenicity and toxicity, and biodegradability. However, current collagen gel formulations fail to recapitulate the range of collagen structures observed in native tissues, presenting a significant challenge in achieving the full potential of collagen-based biomaterials. Collagen fiber structure can be manipulated in vitro through mechanical forces, environmental factors, or thermal mechanisms. Here, we describe a new ultrasound-based fabrication technology that exploits the ability of ultrasound to generate localized mechanical forces to control the collagen fiber microstructure non-invasively. The results indicate that exposing soluble collagen to ultrasound (7.8 or 8.8 ​MHz; 3.2–10 ​W/cm2) during hydrogel formation leads to local variations in collagen fiber structure and organization that support increased levels of cell migration. Furthermore, multiphoton imaging revealed increased cell-mediated collagen remodeling of ultrasound-exposed but not sham-exposed hydrogels, including formation of multicellular aggregates, collagen fiber bundle contraction, and increased binding of collagen hybridizing peptides. Skin explant ​cultures obtained from diabetic mice showed similar enhancement of cell-mediated remodeling of ultrasound-exposed but not sham-exposed collagen hydrogels. Using the mechanical forces associated with ultrasound to induce local changes in collagen fibril structure and organization ​to functionalize native biomaterials is a promising non-invasive and non-toxic technology for tissue engineering and regenerative medicine.

A method for preserving and visualizing the three-dimensional structure of the mouse zonule

Mechanical failure of the ciliary zonule characterizes several ocular and systemic diseases. The mouse has emerged as a useful model system to investigate the composition and structure/function relationships of the zonule. However, visualizing the organization of the diaphanous fibers that comprise the zonule is technically challenging because the fibers do not take up conventional histological stains and are disrupted easily during processing. Here, we describe a simple method for maintaining physiological pressure within the mouse eye during fixation, and a gel-embedding technique for stabilizing the zonular fibers during subsequent tissue processing and imaging steps. This approach facilitates quantitative measurements of fiber number and cross-sectional dimensions and will allow the effects of targeted disruption of zonule components to be assessed systematically.

DDR2 controls breast tumor stiffness and metastasis by regulating integrin mediated mechanotransduction in CAFs.

Biomechanical changes in the tumor microenvironment influence tumor progression and metastases. Collagen content and fiber organization within the tumor stroma are major contributors to biomechanical changes (e., tumor stiffness) and correlated with tumor aggressiveness and outcome. What signals and in what cells control collagen organization within the tumors, and how, is not fully understood. We show in mouse breast tumors that the action of the collagen receptor DDR2 in CAFs controls tumor stiffness by reorganizing collagen fibers specifically at the tumor-stromal boundary. These changes were associated with lung metastases. The action of DDR2 in mouse and human CAFs, and tumors in vivo, was found to influence mechanotransduction by controlling full collagen-binding integrin activation via Rap1-mediated Talin1 and Kindlin2 recruitment. The action of DDR2 in tumor CAFs is thus critical for remodeling collagen fibers at the tumor-stromal boundary to generate a physically permissive tumor microenvironment for tumor cell invasion and metastases.

P51-F Quantitative electromyography: Normative data in paraspinal muscles

Objectives Paraspinal muscle may be affected in neuromuscular disorders, and EMG may be crucial in the diagnosis. The aim of the study was to develop normative data at different spinal segments. Methods Sixty-five healthy subjects (49% men, 51% women) aged 21–82 years were studied. In each subject, 16–92 motor unit potentials (MUPs) were recorded bilaterally at C7, Th10 and L5 segments using concentric needle electrodes. At each segment, average MUP amplitudes, durations, incidence of polyphasic potentials, and the amplitude at maximal voluntary contraction (MVC) were measured. The incidence of spontaneous activity was examined. Results There was no relationship between the EMG parameters and age. The mean MUP duration and amplitude, and the amplitude at MVC increased caudally (ANOVA, P Conclusions In contrast to limb-muscles EMG parameters were unrelated to the age of the subject. The gradient of EMG parameters caudally from C7 suggesting larger motor units at more caudal segments. The proportion of polyphasic potentials may be due to the organization of muscle fibers. The incidence of fibrillation activity was similar to limb muscles. Significance The study describes normative data for EMG of paraspinal muscles according to the examined spinal segment. These data may be useful in quantitative EMG examination of paraspinal muscle in neuromuscular disease.

Characterization of surface orientation and tenderness of sous vide processed edible offal and psoas muscle from Dohne Merino sheep

The microstructural orientation and tenderness of edible offal and the psoas minor muscle from Dohne Merino sheep were compared in this study. Following evisceration and cleaning, fresh meat samples were collected from slaughtered lambs and ewes of different age groups and thermally processed to varying degrees of doneness using sous vide apparatus. A scanning electron microscope (SEM) was used for the characterization of the surface orientation and elemental composition. The results for tenderness showed that edible offal with anisotropic orientation had higher Warner-Bratzler shear force (WBSF) values, but those with some degrees of uniformity (isotropicity) in their surface orientation were comparatively tender. The rumen and trachea from the lambs, six-tooth and eight-tooth sheep at 10 - 12, 23 - 36 and 28 - 48 months old at slaughter had the highest WBSF values among all the meat samples. Our findings showed that edible offal from the lambs with isotropic orientation was more tender and richer in protein than those from ewes. Also, processing the psoas minor muscle and edible offal with sous vide at 100 oC in the shortest cooking time of 30 minutes had the greatest effect on tenderness of the meat samples. In descending order, the psoas minor muscle, liver and tongue from the lambs were found to be inherently more proteinaceous than other meat samples. The differences observed in the tenderness and protein contents of the edible offal and the psoas muscle could be attributed to variations in the microstructural organization of the muscle fibres in Dohne Merino and to their response to thermal processing. Keywords: anisotropy, doneness, isotropy, meat processing, Merino, scanning electron microscopy

Second harmonic generation microscopy of early embryonic mouse hearts.

The understanding of biomechanical regulation of early heart development in genetic mouse models can contribute to improved management of congenital cardiovascular defects and embryonic cardiac failures in humans. The extracellular matrix (ECM), and particularly fibrillar collagen, are central to heart biomechanics, regulating tissue strength, elasticity and contractility. Here, we explore second harmonic generation (SHG) microscopy for visualization of establishing cardiac fibers such as collagen in mouse embryos through the earliest stages of development. We detected significant increase in SHG positive fibrillar content and organization over the first 24 hours after initiation of contractions. SHG microscopy revealed regions of higher fibrillar organization in regions of higher contractility and reduced fibrillar content and organization in mouse Mlc2a model with cardiac contractility defect, suggesting regulatory role of mechanical load in production and organization of structural fibers from the earliest stages. Simultaneous volumetric SHG and two-photon excitation microscopy of vital fluorescent reporter EGFP in the heart was demonstrated. In summary, these data set SHG microscopy as a valuable non-bias imaging tool to investigate mouse embryonic cardiogenesis and biomechanics.

Vitamin D3 constrains estrogen\u2019s effects and influences mammary epithelial organization in 3D cultures

Vitamin D3 (vitD3) and its active metabolite, calcitriol (1,25-(OH)2D3), affect multiple tissue types by interacting with the vitamin D receptor (VDR). Although vitD3 deficiency has been correlated with increased incidence of breast cancer and less favorable outcomes, randomized clinical trials have yet to provide conclusive evidence on the efficacy of vitD3 in preventing or treating breast cancer. Additionally, experimental studies are needed to assess the biological plausibility of these outcomes. The mammary gland of VDR KO mice shows a florid phenotype revealing alterations of developmental processes that are largely regulated by mammotropic hormones. However, most research conducted on vitD3’s effects used 2D cell cultures and supra-physiological doses of vitD3, conditions that spare the microenvironment in which morphogenesis takes place. We investigated the role of vitD3 in mammary epithelial morphogenesis using two 3D culture models. VitD3 interfered with estrogen’s actions on T47D human breast cancer cells in 3D differently at different doses, and recapitulated what is observed in vivo. Also, vitD3 can act autonomously and affected the organization of estrogen-insensitive MCF10A cells in 3D collagen matrix by influencing collagen fiber organization. Thus, vitD3 modulates mammary tissue organization independent of its effects on cell proliferation.

Imaging Collagen Properties in the Uterosacral Ligaments of Women With Pelvic Organ Prolapse Using Spatial Light Interference Microscopy (SLIM)

We studied collagen fiber organization in tissue affected by pelvic organ prolapse (POP) and compared it to asymptomatic controls. Both the control and POP tissue biopsies were prepared and measured by a highly sensitive quantitative phase imaging (QPI) system, called spatial light inference microscopy (SLIM). Combined with automatic image processing, this modality provides quantitative, high-throughput assessment of fiber morphology. We found the fiber orientation in prolapsed specimens is less homogeneous, indicating an abnormal organization of collagen in the extracellular matrix (ECM).