Arrangement Of Collagen Fibrils Research Articles

Comparison of the recovery characteristics for canine corneal ulcer treated with corneoconjunctival transposition or conjunctival autografts

Corneal ulceration induced by Staphylococcus pseudintermedius (S. pseudintermedius) is a common clinical eye disease. Antibiotics combined with corneoconjunctival transposition (CCT) or conjunctival autografts (CA) are often used, but the recovery characteristics are still unknown. In this experiment, canine corneal ulcer models induced by S. pseudintermedius and treated with levofloxacin eye drops (LED) were created. The models were used to compare the recovery characteristics of CCT and CA, combined with LED, by clinical observation, histopathology, and cytokine expression detected by qRT-PCR analysis. The results showed that the ulcerative cornea with only LED treatment perforated after 48 h. The mRNA expression of TLR2, IL-1β, IL-6, IL-8, and TNF-α genes was significantly elevated on 14, 28, and 35 days after the surgery compared to normal (p < 0.01). On day 42, the inflammatory damage had resolved, but the corneal transparency and arrangement of collagen fibrils in the CCT group were higher than those in the CA group. The mRNA expression of EGF, FGF, TGF-β1 and VEGF genes increased significantly (p < 0.01), mostly until day 42, proving that CCT and CA surgery contributed to the corneal recovery, and relieved the inflammatory reaction, with the elimination of corneal cicatrices needing a period of reconstruction. Therefore, this study has provided, for the first time, the method for establishing a canine corneal ulcer model induced by S. pseudintermedius. More importantly, the recovery of canine ulcerative corneas with CCT or CA surgery is reported for the first time.

Investigating cartilage-related diseases by polarization-resolved second harmonic generation (P-SHG) imaging.

Establishing quantitative parameters for differentiating between healthy and diseased cartilage tissues by examining collagen fibril degradation patterns facilitates the understanding of tissue characteristics during disease progression. These findings could also complement existing clinical methods used to diagnose cartilage-related diseases. In this study, cartilage samples from normal, osteoarthritis (OA), and rheumatoid arthritis (RA) tissues were prepared and analyzed using polarization-resolved second harmonic generation (P-SHG) imaging and quantitative image texture analysis. The enhanced molecular contrast obtained from this approach is expected to aid in distinguishing between healthy and diseased cartilage tissues. P-SHG image analysis revealed distinct parameters in the cartilage samples, reflecting variations in collagen fibril arrangement and organization across different pathological states. Normal tissues exhibited distinct χ33/χ31 values compared with those of OA and RA, indicating collagen type transition and cartilage erosion with chondrocyte swelling, respectively. Compared with those of normal tissues, OA samples demonstrated a higher degree of linear polarization, suggesting increased tissue birefringence due to the deposition of type-I collagen in the extracellular matrix. The distribution of the planar orientation of collagen fibrils revealed a more directional orientation in the OA samples, associated with increased type-I collagen, while the RA samples exhibited a heterogeneous molecular orientation. This study revealed that the imaging technique, the quantitative analysis of the images, and the derived parameters presented in this study could be used as a reference for disease diagnostics, providing a clear understanding of collagen fibril degradation in cartilage.

Structural control of corneal transparency, refractive power and dynamics.

The cornea needs to be transparent to visible light and precisely curved to provide the correct refractive power. Both properties are governed by its structure. Corneal transparency arises from constructive interference of visible light due to the relatively ordered arrangement of collagen fibrils in the corneal stroma. The arrangement is controlled by the negatively charged proteoglycans surrounding the fibrils. Small changes in fibril organisation can be tolerated but larger changes cause light scattering. Corneal keratocytes do not scatter light because their refractive index matches that of the surrounding matrix. When activated, however, they become fibroblasts that have a lower refractive index. Modelling shows that this change in refractive index significantly increases light scatter. At the microscopic level, the corneal stroma has a lamellar structure, the parallel collagen fibrils within each lamella making a large angle with those of adjacent lamellae. X-ray scattering has shown that the lamellae have preferred orientations in the human cornea: inferior-superior and nasal-temporal in the central cornea and circumferential at the limbus. The directions at the centre of the cornea may help withstand the pull of the extraocular muscles whereas the pseudo-circular arrangement at the limbus supports the change in curvature between the cornea and sclera. Elastic fibres are also present; in the limbus they contain fibrillin microfibrils surrounding an elastin core, whereas at the centre of the cornea, they exist as thin bundles of fibrillin-rich microfibrils. We present a model based on the structure described above that may explain how the cornea withstands repeated pressure changes due to the ocular pulse.

Effective investigation of murine femoral bone development utilizing correlative light and electron microscopy (CLEM)

BackgroundFor effective investigation of the developing structure and chemistry of bone, comprehensive studies including compositional analysis can be achieved through the gradual observation from the micro- to nanometer scale via correlative light and electron microscopy (CLEM). This technique is particularly useful considering the complex hierarchical arrangement of bioapatite and collagen fibrils which may vary according to specific bone tissue types (i.e., lamellar bone and woven bone) and different growth stages. Scanning electron microscopy (SEM) accompanied with the attachment of the scanning transmission electron microscopy (STEM) detector, referred to as the STEM-in-SEM can be utilized to produce high contrast images from materials composed of light elements, and efficiently allows the selection of suitable accelerating voltage for energy-dispersive spectroscopy (EDS). This study aims to emphasize the efficacy of CLEM techniques through applying STEM-in-SEM and EDS analyses, and its application to comparative murine bone investigation in differing ontogenetic stages.FindingsWe have designed a new grid-holder which can be used for both light and electron microscopy, and we presented an imaging technique for TEM specimens via reflective light microscopy (RLM). For performing CLEM, ultra-thin-sections (UTS) prepared from the femoral bones of 1- to 16-week old of Sprague-Dawley (SD) rats provided light and electron micrographs that can be correlated based on the regions of interest (ROIs). STEM-in-SEM micrographs revealed information not attainable by secondary electron (SE) and back-scattered electron (BSE) micrographs. In addition, for analyzing chemical variation according to growth and development of femoral bones from 1- to 16-week-old rats, comparative chemical analysis was performed through STEM-in-SEM EDS with two reference materials.ConclusionHerein, from femoral bones of SD rats, we have confirmed the rapid chemical and structural variations within the first 8 weeks after birth. STEM-in-SEM micrographs revealed the bone development process of the early stage porous bone matrix subsequently being filled with collagen fibrils and bioapatite. In addition, chemical analysis for carbon and oxygen showed the ratios of inorganic to organic phases according to growth and progress in bone mineralization. As a result, we were able to postulate the growth mechanism of murine femoral bone in the neonatal stages of development. We also anticipate that our CLEM techniques can be further utilized for more thorough investigation of bone structure and chemistry in diverse scales.

Sensitivity of the amide I band to matrix manipulation in bone: a Raman micro-spectroscopy and spatially offset Raman spectroscopy study.

The fracture resistance of bone arises from the hierarchical arrangement of minerals, collagen fibrils (i.e., cross-linked triple helices of α1 and α2 collagen I chains), non-collagenous proteins, and water. Raman spectroscopy (RS) is not only sensitive to the relative fractions of these constituents, but also to the secondary structure of bone proteins. To assess the ability of RS to detect differences in the protein structure, we quantified the effect of sequentially autoclaving (AC) human cortical bone at 100 °C (∼34.47 kPa) and then at 120 °C (∼117.21 kPa) on the amide I band using a commercial Raman micro-spectroscopy (μRS) instrument and custom spatially offset RS (SORS) instrument in which rings of collection fiber optics are offset from the central excitation fiber optics within a hand-held, cylindrical probe. Being clinically viable, measurements by SORS involved collecting Raman spectra of cadaveric femur mid-shafts (5 male & 5 female donors) through layers of a tissue mimic. Otherwise, μRS and SORS measurements were acquired directly from each bone. AC-related changes in the helical status of collagen I were assessed using amide I sub-peak ratios (intensity, I, at ∼1670 cm-1 relative to intensities at ∼1610 cm-1 and ∼1640 cm-1). The autoclaving manipulation significantly decreased the selected amide I sub-peak ratios as well as shifted peaks at ∼1605 cm-1 (μRS), ∼1636 cm-1 (SORS) and ∼1667 cm-1 in both μRS and SORS. Compared to μRS, SORS detected more significant differences in the amide I sub-peak ratios when the fiber optic probe was directly applied to bone. SORS also detected AC-related decreases in I1670/I1610 and I1670/I1640 when spectra were acquired through layers of the tissue mimic with a thickness ≤2 mm by the 7 mm offset ring, but not with the 5 mm or 6 mm offset ring. Overall, the SORS instrument was more sensitive than the conventional μRS instrument to pressure- and temperature-related changes in the organic matrix that affect the fracture resistance of bone, but SORS analysis of the amide I band is limited to an overlying thickness layer of 2 mm.

A mouse model of brittle cornea syndrome type 2

AbstractPurpose: Brittle Cornea Syndrome (BCS) is a rare disease that affects 1 in 1 000 000 people world wide. With a reduction in cornea thickness and therefore strength sufferers have an increase in loss of vision through injury. Our lab has previously published the first mouse model of BCS Type 1(1) and are currently characterizing a novel model of BCS type 2 in order to elucidate the underlying mechanisms causing this disease.Methods: Mice carrying a 7 bp deletion which causes a frameshift and premature stop codon within the causative gene Prdm5 have been extensively phenotyped using Optical Coherence Tomography (OCT), Transmission Electron Microscopy (TEM) and Histology.Results: These mice show a reduced central corneal thickness by OCT from as early as 1 month of age. TEM and histological analysis indicates that this reduction in thickness may be due to the dysregulation of collagen assembly – as stromal collagen fibre diameter is significantly reduced.Conclusions: The structure of the stroma is crucial to corneal function: mechanical strength and transparency are achieved by the intricate arrangement of collagen fibrils, synthesized by keratocytes. Although it is known that the corneal stroma has a unique composition in which the ratio of Type I to Type V collagen is very different to anywhere else in the body, it remains unclear how the synthesis and maintenance of this specialized extracellular matrix by keratocytes is controlled. Characterization of the nature and extent of developmental defects in the cornea of this mouse model will be important to devise future therapeutic interventions.Reference1. Stanton, C. M., Findlay, A. S., Drake, C., Mustafa, M. Z., Gautier, P., McKie, L., Jackson, I. J., &amp; Vitart, V. (2021). A mouse model of brittle cornea syndrome caused by mutation in Zfp469. Disease models &amp; mechanisms, 14(9), dmm049175. https://doi.org/10.1242/dmm.049175

Structure and Strength of Bovine and Equine Amniotic Membrane.

Simple SummaryThin, strong scaffold materials are needed for surgical applications. There is a limited selection of available materials and new materials are required. Amnionic membrane from cattle and horses were investigated for this purpose. The structure of these materials was characterized with synchrotron techniques and the strength was measured. A possible relationship between the structure and strength was identified. These amnion materials from animal sources are strong, thin, and elastic materials, although weaker than some other collagen tissues. They may be suitable for use in surgery as an alternative to material from human donors.Thin, strong scaffold materials are needed for surgical applications. New materials are required, particularly those readily available, such as from non-human sources. Bovine amniotic membrane (antepartum) and equine amniotic membrane (postpartum) were characterized with tear and tensile tests. The structural arrangement of the collagen fibrils was determined by small-angle X-ray scattering, scanning electron microscopy, and ultrasonic imaging. Bovine amnion had a thickness-normalized tear strength of 12.6 (3.8) N/mm, while equine amnion was 14.8 (5.3) N/mm. SAXS analysis of the collagen fibril arrangement yielded an orientation index of 0.587 (0.06) and 0.681 (0.05) for bovine and equine, respectively. This may indicate a relationship between more highly aligned collagen fibrils and greater strength, as seen in other materials. Amnion from bovine or equine sources are strong, thin, elastic materials, although weaker than other collagen tissue materials commonly used, that may find application in surgery as an alternative to material from human donors.

Corneal transparency and scleral opacity arises from the nanoarchitecture of the constituent collagen fibrils.

While human scleral and corneal tissues possess similar structural morphology of long parallel cylindrical collagen fibrils, their optical characteristics are markedly different. Using pseudospectral time-domain (PSTD) simulations of Maxwell's equations, we model light propagation through realistic representations of scleral and corneal nanoarchitecture and analyze the transmittance and spatial correlation in the near field. Our simulation results provide differing predictions for scleral opacity and corneal transparency across the vacuum ultraviolet to the mid-infrared spectral region in agreement with experimental data. The simulations reveal that the differences in optical transparency between these tissues arise through differences in light scattering emanating from the specific nanoscale arrangement and polydispersity of the constituent collagen fibrils.

An ultrastructural 3D reconstruction method for observing the arrangement of collagen fibrils and proteoglycans in the human aortic wall under mechanical load

An insight into changes of soft biological tissue ultrastructures under loading conditions is essential to understand their response to mechanical stimuli. Therefore, this study offers an approach to investigate the arrangement of collagen fibrils and proteoglycans (PGs), which are located within the mechanically loaded aortic wall. The human aortic samples were either fixed directly with glutaraldehyde in the load-free state or subjected to a planar biaxial extension test prior to fixation. The aortic ultrastructure was recorded using electron tomography. Collagen fibrils and PGs were segmented using convolutional neural networks, particularly the ESPNet model. The 3D ultrastructural reconstructions revealed a complex organization of collagen fibrils and PGs. In particular, we observed that not all PGs are attached to the collagen fibrils, but some fill the spaces between the fibrils with a clear distance to the collagen. The complex organization cannot be fully captured or can be severely misinterpreted in 2D. The approach developed opens up practical possibilities, including the quantification of the spatial relationship between collagen fibrils and PGs as a function of the mechanical load. Such quantification can also be used to compare tissues under different conditions, e.g., healthy and diseased, to improve or develop new material models. Statement of significanceThe developed approach enables the 3D reconstruction of collagen fibrils and proteoglycans as they are embedded in the loaded human aortic wall. This methodological pipeline comprises the knowledge of arterial mechanics, imaging with transmission electron microscopy and electron tomography, segmentation of 3D image data sets with convolutional neural networks and finally offers a unique insight into the ultrastructural changes in the aortic tissue caused by mechanical stimuli.

Decorin regulates collagen fibrillogenesis during corneal wound healing in mouse in vivo

A characteristic rigid spatial arrangement of collagen fibrils in the stroma is critical for corneal transparency. This unique organization of collagen fibrils in corneal stroma can be impacted by the presence and interactions of proteoglycans and extracellular matrix (ECM) proteins in a corneal microenvironment. Earlier studies revealed that decorin, a leucine-rich proteoglycan in stroma, regulates keratocyte-collagen matrix assembly and wound healing in the cornea. This study investigated the role of decorin in the regulation of stromal fibrillogenesis and corneal transparency in vivo employing a loss-of-function genetic approach using decorin null (dcn−/−) and wild type (dcn+/+) mice and a standard alkali-injury model. A time-dependent ocular examinations with Slit lamp microscope in live animals assessed corneal clarity, haze, and neovascularization levels in normal and injured eyes. Morphometric changes in normal and injured dcn+/+ and dcn−/− corneas, post-euthanasia, were analyzed with Masson's Trichrome and Periodic Acid-Schiff (PAS) histology evaluations. The ultrastructure changes in all corneas were investigated with transmission electron microscopy (TEM). Injury to eye produced clinically relevant corneal haze and neovascularization in dcn−/− and dcn+/+ mice while corneas of uninjured eyes remained clear and avascular. A clinically significant haze and neovascularization appeared in injured dcn−/- corneas compared to the dcn+/+ corneas at day 21 post-injury and not at early tested times. Histological examinations revealed noticeably abnormal morphology and compromised collagen levels in injured dcn−/− corneas compared to the injured/normal dcn+/+ and uninjured dcn−/− corneas. TEM analysis exhibited remarkably uneven collagen fibrils size and distribution in the stroma with asymmetrical organization and loose packing in injured dcn−/− corneas than injured/normal dcn+/+ and uninjured dcn−/− corneas. The minimum and maximum inter-fibril distances were markedly irregular in injured dcn−/− corneas compared to all other corneas. Together, results of clinical, histological, and ultrastructural investigations in a genetic knockout model suggested that decorin influenced stromal fibrillogenesis and transparency in healing cornea.

Histological and ultrastructural characterisation as minimal criteria for assessing the success of the decellularisation protocols for tissue engineering applications

There is a great demand of suitable biomaterials for tissue engineering applications in regenerative medicine. Decellularized extracellular matrix scaffolds derived from different organs and tissues have been successfully applied for human use as therapeutic agents in many tissue defects. The aim of the present study was to establish histological and ultrastructural characterisation as minimal criteria for assessing the suitability of decellularised bioscaffolds in tissue engineering applications. The study was performed on bovine omentum processed by a natural bovine bile based decellularisation protocol. Histological analysis and scanning electron microscopy examinations were performed and compared with an established decellularised bovine pericardial scaffold. Gross examination revealed that the decellularisation process did not alter the shape of the omentum. Histological examination confirmed the preservation of the collagenous fibres without any observable cell nuclei following decellularisation. Further, scanning electron microscopy examination demonstrated the arrangement and ultrastructure of collagen fibrils in the bioscaffolds. Our results suggest that histological and ultrastructural characterisation provide preliminary data on determining the biocompatibility of scaffolds before planning in vivo studies in tissue engineering applications.

Analysis and simulation of fracture behavior in naturally occurring Bouligand structures

Through natural selection processes, refined biological materials could be created that adapt to various environments and exhibit specific functions. Such materials include typical Bouligand structures that can be widely observed in marine creatures that have hard shells. Consisting of a helicoidal arrangement of aligned fibrils, layered single-twisted Bouligand-type structures (SBS) display exceptional fracture and damage resistance. A much more primitive and rarer type of this formation, the double-twisted Bouligand-type structures (DBS), has been discovered in ancient fish scales, and this architecture could provide added rigidity and significantly contribute to toughness when facing fracture risk. In this work, we describe a computational modeling approach to investigating fracture behaviors and toughening mechanisms in Bouligand structures. To achieve qualitative insights into the fracture behaviors of DBS and SBS, we applied these two configurations, which were identified from corresponding biological materials, to analyze load-displacement responses during single edge notched (SEN) tensile testing; the toughening mechanism is also discussed further. The results clearly show that the arrangement of helix fibrils and interlaminar properties play a major role in the resulting fracture behaviors of Bouligand architectures. This is of interest for the future design of engineering materials and structures that require composites with enhanced toughness, and could deepen our understanding of the structure-property relationship of Bouligand-type structures in bionic design. Statement of significanceIn this work, a novel numerical modeling approach based on the extended finite element method (XFEM) has been established to evaluate the fracture behavior of a naturally-occurring Bouligand-type helicoidal structure subjected to the single edge notched (SEN) tensile loading. The roles of the biological features (i.e., layered arrangement of collagen fibrils and interbundle fibrils) on the fracture resistance and toughening mechanism of the Bouligand-type structures have been uncovered and analyzed quantitatively. This is of interest for future design of engineering materials and structures that require composites with enhanced toughness, and can deepen the understanding of the structure-property relationship of the Bouligand-type structure in bionic design.

Proteoglycan and collagen contribution to the strain-rate-dependent mechanical behaviour of knee and shoulder cartilage

The contribution of the proteoglycan to the strain-rate-dependent mechanical behaviour of cartilage tissues has been suggested to decrease with an increase in the strain-rate. On the other hand, the contribution from the collagen network has been suggested to increase as the strain-rate increases. These conclusions are drawn mainly based on numerical studies conducted on high-load-bearing knee cartilage tissues, while experimental evidence of these behaviours have not been demonstrated previously. Further, in contrast to the reported findings on high-load bearing knee cartilage, our previous study on the low-load-bearing kangaroo shoulder cartilage indicated that proteoglycan and collagen contribution remained steady as the strain-rate increases. Therefore, in the present study, we experimentally investigate the contribution of proteoglycan and collagen network to the strain-rate-dependent behaviour of the kangaroo knee cartilage, and plausible reasons for the differences observed in relation to the kangaroo shoulder cartilage. Firstly, in order to quantify the contribution of proteoglycans and collagen network, the indentation testings on normal, proteoglycan, and collagen-degraded kangaroo knee cartilage were conducted at different strain-rates. Then, structural and compositional differences between the kangaroo knee and shoulder cartilage were assessed qualitatively through polarised light microscopy (PLM) imaging and histological staining. Identified differences in the collagen architecture and proteoglycan composition were incorporated in a fibril-reinforced porohyperelastic Finite Element (FE) model with the objective of explaining the mechanisms underlying differences observed between the two tissues. Experimental results on knee cartilage indicated that when the strain-rate increases, proteoglycan contribution decreases while collagen contribution increases, where statistically significant differences were identified at each strain-rate (p<0.05). PLM images revealed a sizable deep zone in the kangaroo knee cartilage where collagen fibrils were oriented perpendicular to the subchondral bone. On the other hand, no such apparent deep zone was observed in the shoulder cartilage. FE model confirmed that the biomechanical differences observed in the knee and shoulder cartilage are due to the differences in the collagen fibril arrangement in the deep zone. From these results, it can be concluded that in high-load-bearing cartilage tissues, the collagen network in the deep zone assists in increasing the stiffness of tissue with strain-rate and plays a significant role in supporting transient loads. This, in turn, helps protect the solid matrix against large distortions and strains at the subchondral junction, pointing to the importance of the collagen network in deep zone in assisting high-load-bearing cartilage tissues.

Differences in MPS I and MPS II Disease Manifestations.

Mucopolysaccharidosis (MPS) type I and II are two closely related lysosomal storage diseases associated with disrupted glycosaminoglycan catabolism. In MPS II, the first step of degradation of heparan sulfate (HS) and dermatan sulfate (DS) is blocked by a deficiency in the lysosomal enzyme iduronate 2-sulfatase (IDS), while, in MPS I, blockage of the second step is caused by a deficiency in iduronidase (IDUA). The subsequent accumulation of HS and DS causes lysosomal hypertrophy and an increase in the number of lysosomes in cells, and impacts cellular functions, like cell adhesion, endocytosis, intracellular trafficking of different molecules, intracellular ionic balance, and inflammation. Characteristic phenotypical manifestations of both MPS I and II include skeletal disease, reflected in short stature, inguinal and umbilical hernias, hydrocephalus, hearing loss, coarse facial features, protruded abdomen with hepatosplenomegaly, and neurological involvement with varying functional concerns. However, a few manifestations are disease-specific, including corneal clouding in MPS I, epidermal manifestations in MPS II, and differences in the severity and nature of behavioral concerns. These phenotypic differences appear to be related to different ratios between DS and HS, and their sulfation levels. MPS I is characterized by higher DS/HS levels and lower sulfation levels, while HS levels dominate over DS levels in MPS II and sulfation levels are higher. The high presence of DS in the cornea and its involvement in the arrangement of collagen fibrils potentially causes corneal clouding to be prevalent in MPS I, but not in MPS II. The differences in neurological involvement may be due to the increased HS levels in MPS II, because of the involvement of HS in neuronal development. Current treatment options for patients with MPS II are often restricted to enzyme replacement therapy (ERT). While ERT has beneficial effects on respiratory and cardiopulmonary function and extends the lifespan of the patients, it does not significantly affect CNS manifestations, probably because the enzyme cannot pass the blood–brain barrier at sufficient levels. Many experimental therapies, therefore, aim at delivery of IDS to the CNS in an attempt to prevent neurocognitive decline in the patients.

Ultrastructural Alterations of Grafted Corneal Buttons: The Anatomic Basis for Stromal Peeling Along a Natural Plane of Separation

To examine the ultrastructure of the natural plane of separation in grafted corneas and evaluate the outcomes of stromal peeling. Interventional case series. In this multicenter study, stromal peeling was attempted in 96 consecutive eyes with unsatisfactory vision following penetrating keratoplasty (PK) for keratoconus (n=79), herpetic keratitis (n=11), and granular dystrophy (n=6). Stromal exchange was performed by (1) 9 mm partial-thickness trephination; (2) creation of a corneal flap across the PK wound; (3) opening of the stromal component of the PK wound until a smooth, translucent natural plane was identified; (4) severing the attachment of the PK scar; (5) stromal peeling along the identified plane; and (6) suturing of donor lamella. Grafted corneas from cases that mandated conversion to PK were processed for transmission electron microscopy. The natural plane of separation was identified in all cases. Stromal exchange was successfully completed in 84 cases (87.5%). Snellen visual acuity ≥20/40 and ≥20/25 was reached in 93% and 72% of cases at 3 years (n=49) and 86% and 62% at 4 years (n=21) postoperatively. Mean endothelial cell loss at 1 year was 6.6% ± 9.5%. Stromal peeling occurred along a plane lined with a continuous layer of keratocytes separating pre-Descemet membrane (DM) stroma, DM, and endothelium from the anterior stroma. Pre-DM stroma was made of poorly organized lamellae containing widely spaced, randomly arranged collagen fibrils. Ultrastructural alterations in the stromal microarchitecture of grafted corneas provide evidence of a natural plane of separation identified intraoperatively. Stromal peeling can be successfully performed in post-PK eyes with various stromal pathology.

A Theoretical Study on the Mechanical Significance of Mineralized Collagen Fibril Orientation in Osteonal Lamellar Bone

In this study, the effect of orientation of mineralized collagen fibrils on bone mechanical properties relating to bone anisotropy was studied using data obtained from rehydrated lamellar bone samples. The dehydration-rehydration based and experimentally determined contraction, observed in orientations parallel and perpendicular to the osteonal axis was used to calculate bone anisotropy. The sublamellar bone model, with the layered mineralized collagen fibrils rotating at 5° was used. Following this model, the mineralized collagen layers were transformed at 5° relative to the orthogonal axes using a transform matrix. With dehydration, fibril diameter was reduced towards the mineral, forming contraction vectors. The x, y and z intercepts for these vectors were then calculated to give the u, v and w displacements, which gave anisotropy ratios ranging from 0.15266 to 6.55054. Compared with the experimental nanoindentation findings in the literature, there may be an indication of a correlation with the results of sublamellar arrangement at 20° angles. As the lateral indentation used in the anisotropy experiments may involve varying amounts of u and v displacements, the aspect angle of lateral indentation was evaluated in relation to the structural features of the model. This evaluation indicated the larger contribution of v displacement and thus relatively much smaller contribution of u displacement to lateral contraction. These findings indicate the significant effect of the mineralized collagen fibril arrangement on bone anisotropy.

Extracellular Matrix Deposition and Remodeling after Corneal Alkali Burn in Mice.

Corneal transparency relies on the precise arrangement and orientation of collagen fibrils, made of mostly Type I and V collagen fibrils and proteoglycans (PGs). PGs are essential for correct collagen fibrillogenesis and maintaining corneal homeostasis. We investigated the spatial and temporal distribution of glycosaminoglycans (GAGs) and PGs after a chemical injury. The chemical composition of chondroitin sulfate (CS)/dermatan sulfate (DS) and heparan sulfate (HS) were characterized in mouse corneas 5 and 14 days after alkali burn (AB), and compared to uninjured corneas. The expression profile and corneal distribution of CS/DSPGs and keratan sulfate (KS) PGs were also analyzed. We found a significant overall increase in CS after AB, with an increase in sulfated forms of CS and a decrease in lesser sulfated forms of CS. Expression of the CSPGs biglycan and versican was increased after AB, while decorin expression was decreased. We also found an increase in KS expression 14 days after AB, with an increase in lumican and mimecan expression, and a decrease in keratocan expression. No significant changes in HS composition were noted after AB. Taken together, our study reveals significant changes in the composition of the extracellular matrix following a corneal chemical injury.

Three-Dimensional Bioprinting for Tissue Engineering and Regenerative Medicine in Down Under: 2020 Australian Workshop Summary.

Three-Dimensional Bioprinting for Tissue Engineering and Regenerative Medicine in Down Under: 2020 Australian Workshop Summary.

Constitutive Modeling of Corneal Tissue: Influence of Three-Dimensional Collagen Fiber Microstructure.

The cornea, the transparent tissue in the front of the eye, along with the sclera, plays a vital role in protecting the inner structures of the eyeball. The precise shape and mechanical strength of this tissue are mostly determined by the unique microstructure of its extracellular matrix. A clear picture of the 3D arrangement of collagen fibrils within the corneal extracellular matrix has recently been obtained from the secondary harmonic generation images. However, this important information about the through-thickness distribution of collagen fibrils was seldom taken into account in the constitutive modeling of the corneal behavior. This work creates a generalized structure tensor (GST) model to investigate the mechanical influence of collagen fibril through-thickness distribution. It then uses numerical simulations of the corneal mechanical response in inflation experiments to assess the efficacy of the proposed model. A parametric study is also done to investigate the influence of model parameters on numerical predictions. Finally, a brief comparison between the performance of this new constitutive model and a recent angular integration (AI) model from the literature is given.

Effects of oxidation on the structure of collagen fibers of sea cucumber (Apostichopus japonicus) body wall during thermal processing

Thermal treatment during sea cucumber processing might affect the texture of the final product. In the present study, collagen fibers (CFs) extracted from the body wall of sea cucumber Apostichopus japonicus were used to investigate the effects of heating and oxidative conditions on CFs structure. Hydroxyl radicals (•OH) were generated in CFs treated at 37 °C and the intensity of the signal was comparable to samples under oxidative conditions at 4 °C. Release of protein and glycosaminoglycan was observed in CFs heat-treated at 37 °C or under oxidative conditions at 4 °C, leading to the conversion of α-helixes into β-sheets, red shift of amide band I, decrease in thermostability, and scattered arrangement of collagen fibrils. The degree of damage in CFs structure is different among groups. In particular, in thermally- and oxidative treated group, macromolecular fragments remarkably degraded over time, 10 kDa proteins were abundantly released, amide bands A and III showed redshift, and maximum denaturation temperature and decomposition temperature were the lowest compared to other groups. The findings discussed herein reveal the structural changes induced by thermal treatment in sea cucumber CFs and provide an explanation of the mechanism from the view of protein oxidation.