Distribution Of Collagen Fibrils Research Articles

Histopathological findings of anterior lens capsule in pediatric cataract.

To investigate the histopathological findings of the anterior lens capsule in pediatric patients who had surgery for cataracts. This study is a prospective interventional study. Anterior capsule tissue samples that were obtained by the anterior capsulotomy method during phacoemulsification surgery were fixed and examined under a transmission electron microscope. Twenty-two eyes of 19 patients who were diagnosed with congenital and juvenile cataracts were included in this study. Five patients had associated systemic diseases, including hydrocephalus, cerebral palsy, prematurity, juvenile myelomonocytic leukemia, and Down's syndrome. Electron microscopic evaluation demonstrated single-layered epithelium under the capsule, degenerated organelles with round-oval and prismatic-oval nuclei, and degenerated mitochondria and heterochromatin-rich nuclei. In the case with cerebral palsy, collagen fibrils of the connective tissue and fibroblast-like cells were observed replacing the epithelium that should be underneath the capsule in both eyes, and there was a disorganized distribution of collagen fibrils and vacuole structures in the cytoplasm of fibroblast-like cells. Similar histopathological findings were found in pediatric cataracts with or without systemic disease except in one cerebral palsy case. The absence of lens epithelium may have been a result of degeneration in this patient, and this can be attributed to the presence of systemic inflammation and gliosis in cerebral palsy. The absence of lens epithelium can play a role in the development of dense subcapsular fibrosis and cataract formation.

Biomimetic grafts from ultrafine fibers for collagenous tissues.

The ligament is the soft tissue that connects bone to bone and, in case of severe injury or rupture, it cannot heal itself mainly because of its poor vascularity and dynamic nature. Tissue engineering carries the potential to restore the injured tissue functions by utilization of scaffolds mimicking the structure of native ligament. Collagen fibrils in the anterior cruciate ligament (ACL) have a diameter ranging from 20 to 300nm, which defines the physical and mechanical properties of the tissue. Also, the ACL tissue exhibited a bimodal distribution of collagen fibrils. Currently, the ability to fabricate scaffolds replicating this structure is a significant challenge. This work aims at i) measuring the diameter of collagens of bovine ACL tissue, ii) investigating the fabrication of sub-100nm fibers, and iii) fabricating aligned scaffolds with bimodal diameter distribution (with two peaks) resembling the healthy ACL structure. It is hypothesized that such scaffolds can be produced by electrospinning polycaprolactone (PCL) solutions. To test the hypothesis, various PCL solutions were formulated in acetone and formic acid in combination with pyridine, and electrospun to generate sub-100nm fibers. Next, this formulation was adjusted to produce nanofibers with a diameter between 100nm and 200nm. Finally, these solutions were combined in the co-electrospinning process, i.e., two-spinneret electrospinning, to fabricate biomimetic scaffolds with a bimodal distribution. Electrospinning of 8% and 15% PCL solutions, respectively, resulted in the production of fibers with diameters below and above 100nm. The combined scaffold exhibited a bimodal distribution of aligned fibers with peaks around 80 and 180nm, thus mimicking the collagen fibrils of healthy ACL tissue. This research is expected to have a society-wide impact because it aims to enhance the health condition and life quality of a wide range of patients.

Modeling the deterioration of the stiffness and of the collagen fibril distribution in a discrete model of the cornea microstructure

We present a stochastic approach to model the mechanical deterioration of the reinforcing microstructure of the human cornea. The fundamental structural micro-components of the stroma, collagen and crosslinks, are assembled deterministically into an elementary trusswork cell, multiply repeated and distorted to form a three-dimensional shell with the shape of a cornea. The spatial orientation of the collagen-like elements of each cell is thus characterized stochastically with a non correlated random angle field, obeying an assigned probability density function, leading to a non-deterministic structural stiffness. It follows that the mechanical response of the model to the action of deterministic forces equivalent to the intraocular pressure is stochastic due to the random spatial orientation of collagen fibers. The deterioration of the mechanical stiffness of the collagen components is described through a scalar variable field, evolving in space and in time, representative of a progressive damage which causes heterogeneity and asymmetric behavior. The damage variable acts in two ways on the global stiffness: (i) by reducing the stiffness of the collagen components; (ii) by modifying the dispersion coefficient of the probability density function. The equilibrium equations of the damaging model are solved at discrete time steps, with a fully explicit solution scheme, by means of the stochastic finite element improved perturbation method. The results show that when the collagen fibril stiffness reduces to 10% of the healthy value, as expected in the case of the vision-impairing condition known as keratoconus, the displacement field due to intraocular pressure is significantly affected in terms of both average and variance distributions. This effect confers a typical conical shape to the cornea. In particular, the analysis shows that high values of the response variances are confined in the keratoconus area, which agrees with a high level of uncertainties due to loss of fibril organization and thickness reduction under pathologic conditions.

Glycosaminoglycans and collagen fibril distribution at various depths of the corneal stroma of normal and CXL treated rats

Corneal collagen cross-linking (CXL) is widely used to treat keratoconus and ecstatic corneal disorders. The present studies were carried out to investigate the distribution of glycosaminoglycans (GAGs) and collagen fibril (CF) at different depths of the normal and CXL treated corneal stroma of four week old rats 7 days after standard CXL application. Ten Wistar rats’ corneas were used for the study. The epithelium of the cornea from the left eye of each rat was removed and treated with standard CXL application using riboflavin and Ultraviolet-A (UVA) (3 mW/cm2 for 30 min). The cornea from the right eye was used as the control cornea. The cornea was removed from the eye and processed for transmission electron microscopy. A bottom mounted Quemesa camera was used to capture digital images and these images were analysed using iTEM software. In the control cornea, the GAGs area size was not significantly different in the anterior, middle, and posterior stroma. In the CXL treated rats the GAGs area size gradually increased from the anterior to the posterior stroma whereas the spacing between the GAGs gradually decreased. There were very large GAGs present in the posterior stroma of the CXL treated rats. When comparing the control and CXL cornea, the GAGs area in the CXL cornea was significantly higher and inter-GAGs-spacing was smaller than in the control cornea. In the control cornea, the collagen fibrils diameter was higher in the anterior stroma and lowest in the posterior stroma. In the CXL treated cornea, the CF diameter and the interfibrillar spacing gradually decreased from the anterior to the posterior stroma. On comparison between the control and the CXL treated cornea, the interfibrillar spacing was significantly smaller in the CXL treated cornea than the control cornea in the anterior, middle, and posterior stroma but there was no difference in the diameter. The CXL treatment significantly increased the GAGs area and decreased the inter-GAGs-spacing, and inter–CF–spacing. This could be due to the gradual decline in the availability of riboflavin, UVA, and oxygen in the middle and posterior stroma. Further studies are required to investigate the role of keratan sulphate and chondroitin sulphate by using monoclonal antibodies with immunogold technique.

Collagen Fibril Diameter Distribution of Sheep Anterior Cruciate Ligament.

The anterior cruciate ligament (ACL) tissue is a soft tissue connecting the femur and tibia at the knee joint and demonstrates a limited capacity for self-regeneration due to its low vascularity. The currently available clinical procedures are unable to fully restore damaged ACL tissue, and tissue engineering can offer options with a potential of restoring the torn/ruptured ACL by using biomimetic constructs that are similar to native tissue in terms of structure, composition, and functions. However, a model substrate to understand how the ACL cells regenerate the injured tissue is still not available. In this study, it is hypothesized that the nanofiber-based model substrate with bimodal and unimodal fiber diameter distributions will mimic the diameter distribution of collagen fibrils seen in healthy and injured sheep ACL, respectively. The aims were to (i) create an ACL injury in a sheep ACL by applying extensional force to rupture the healthy ACL tissue, (ii) measure the collagen fibril diameter distributions of healthy and injured ACL, (iii) fabricate polycaprolactone (PCL) nanofiber-based model constructs using electrospinning with diameter distributions similar to healthy and injured ACL tissue, and (iv) measure mechanical properties of ACL tissue and PCL electrospun constructs. The results showed that the fiber diameter distributions of PCL electrospun constructs and those of the healthy and injured ACL tissues were similar. The novelty in this investigation is that the collagen fibril diameter distribution of healthy and injured sheep ACL tissues was reported for the first time. The study is significant because it aims to create a model construct to solve an important orthopedic-related clinical problem affecting millions of people globally. The model construct fabricated in this work is expected to have an important impact on ACL regeneration efforts.

A numerical model for fibril remodeling in articular cartilage

BackgroundCollagen fibrils of articular cartilage have a distinct organization in mature human knee joints. It seems that a mechanobiological process drives the remodeling of newborn collagen fibrils with maturation. Therefore, the goal of the present study was to develop a collagen fibril remodeling algorithm that describes the unique collagen fibril organization in a 3D knee model. MethodA fibril-reinforced, biphasic cartilage model was used with a cuboid and a 3D human knee joint geometries. An isotropic collagen fibril distribution was assigned to the cartilage at the start of the analysis. Each fibril was rotated towards the direction that resulted in a maximum stretch at each time increment of the loading cycle. ResultsThe resulting pattern for the collagen fibrils was compared with split line patterns of porcine knee joint cartilage and also data published in the literature. Fibrils on the articular surface had a radial pattern towards the geometrical centroid of the tibial and femoral cartilage. In the tibiofemoral contact regions of superficial zone, fibrils were oriented circumferentially and randomly. In the porcine samples, the split-line patterns were similar to those obtained theoretically. Depth-wise organization of fibril network was characterized by fibrils perpendicular to the subchondral bone in the deeper layers, and fibrils parallel to the surface of cartilage in the superficial zone. ConclusionsThe maximum stretch criterion, coupled with a biphasic constitutive model, successfully predicted the collagen fibril organization observed in the articular cartilage throughout the depth and on the articular surface.

Systematic evaluation of three porcine-derived collagen membranes for guided bone regeneration.

Guided bone regeneration is one of the most common surgical treatment modalities performed when an additional alveolar bone is required to stabilize dental implants in partially and fully edentulous patients. The addition of a barrier membrane prevents non-osteogenic tissue invasion into the bone cavity, which is key to the success of guided bone regeneration. Barrier membranes can be broadly classified as non-resorbable or resorbable. In contrast to non-resorbable membranes, resorbable barrier membranes do not require a second surgical procedure for membrane removal. Commercially available resorbable barrier membranes are either synthetically manufactured or derived from xenogeneic collagen. Although collagen barrier membranes have become increasingly popular amongst clinicians, largely due to their superior handling qualities compared to other commercially available barrier membranes, there have been no studies to date that have compared commercially available porcine-derived collagen membranes with respect to surface topography, collagen fibril structure, physical barrier property, and immunogenic composition. This study evaluated three commercially available non-crosslinked porcine-derived collagen membranes (Striate+TM, Bio-Gide® and CreosTM Xenoprotect). Scanning electron microscopy revealed similar collagen fibril distribution on both the rough and smooth sides of the membranes as well as the similar diameters of collagen fibrils. However, D-periodicity of the fibrillar collagen is significantly different among the membranes, with Striate+TM membrane having the closest D-periodicity to native collagen I. This suggests that there is less deformation of collagen during manufacturing process. All collagen membranes showed superior barrier property evidenced by blocking 0.2-16.4 μm beads passing through the membranes. To examine the immunogenic agents in these membranes, we examined the membranes for the presence of DNA and alpha-gal by immunohistochemistry. No alpha-gal or DNA was detected in any membranes. However, using a more sensitive detection method (real-time polymerase chain reaction), a relatively strong DNA signal was detected in Bio-Gide® membrane, but not Striate+TM and CreosTM Xenoprotect membranes. Our study concluded that these membranes are similar but not identical, probably due to the different ages and sources of porcine tissues, as well as different manufacturing processes. We recommend further studies to understand the clinical implications of these findings.

Histopathological findings of anterior lens capsule in paediatric cataract patients

AbstractPurpose: To investigate the histopathological findings of the anterior lens capsule in patients with paediatric cataracts.Methods: This study is a prospective interventional study. Anterior capsule tissue samples which were obtained by anterior capsulotomy method during phaco surgery were fixed in 0.1 M phosphate buffered 2.5% glutaraldehyde at +4°C for 2–4 h and after passing through other stages, thin sections were taken and stained. It was then examined and visualized under HITACHI HT7800 transmission electron microscope.Results: Twenty‐three eyes of 19 patients were included. Three patients had unilateral cataract and remaining 16 had bilateral cataract. Four patients had concomitant systemic disease including hydrocephalus, cerebral palsy, juvenile myelomonocytic leukaemia and Down syndrome. Except for one patient, common findings were single‐layered epithelium under the capsule, degenerated organelles with round‐oval and prismatic‐oval nuclei in some regions, enlarged mitochondria and heterochromatin cells. In the case with cerebral palsy, unlike the normal lens structure, collagen fibrils of the connective tissue and fibroblast‐like cells were observed in the epithelial area that should be located under the capsule, in both eyes. Disorganized distribution of collagen fibrils and vacuole structures in the cytoplasm of fibroblast‐like cells were observed.Conclusions: Similar histopathological findings were found in paediatric cataracts with or without systemic disease. Due to the presence of increased inflammation and gliosis in cerebral palsy, the absence of lens epithelium may have developed as a result of degeneration in this patient. The absence of lens epithelium and inflammation are thought to play a role in the development of dense subcapsular fibrosis.

Periostin modulates extracellular matrix behavior in tendons

Periostin, originally named osteoblast-specific factor 2 (OSF-2) has been identified primarily in collagen rich, biomechanically active tissues where its role has been implicated in mechanisms to maintain the extracellular matrix (ECM), including collagen fibrillogenesis and crosslinking. It is well documented that periostin plays a role in wound healing and scar formation after injury, in part, by promoting cell proliferation, myofibroblast differentiation, and/or collagen fibrillogenesis. Given the significance of periostin in other scar forming models, we hypothesized that periostin will influence Achilles tendon healing by modulating ECM production. Therefore, the objective of this study was to elucidate the effects of periostin during Achilles tendon healing using periostin homozygous (Postn−/−) and heterozygous (Postn+/-) mouse models. A second experiment was included to further examine the influence of periostin on collagen composition and function using intact dorsal tail tendons. Overall, Postn−/− and Postn+/- Achilles tendons exhibited impaired healing as demonstrated by delayed wound closure, increased type III collagen production, decreased cell proliferation, and reduced tensile strength. Periostin ablation also reduced tensile strength and stiffness, and altered collagen fibril distribution in the intact dorsal tail tendons. Achilles tendon outcomes support our hypothesis that periostin influences healing, while tail tendon results indicate that periostin also affects ECM morphology and behavior in mouse tendons.

Особенности ультраструктурных изменений роговичного коллагена экспериментальных животных после уф кросслинкинга роговицы

Purpose. Electron microscopy analysis of collagen fibrils in the corneal stroma of rats after UV cross-linking. Material and methods. Experiments were carried out on rats in 2 groups: 1st – control (intact), 2nd – experiment (UV corneal cross-linking with riboflavin). The observation period is 3, 90 days. The UV irradiation power of the cornea is 3 mW / cm², the duration is 10 minutes. For electron microscopic examination, corneal pieces were fixed in a solution of 2 % glutaraldehyde. Dehydration and pouring were carried out according to the generally accepted method. Ultrathin sections were obtained using the ultramicrotome "Leica EM UC7" (Leica Microsystems) and viewed in the electron microscope JEM-1011 (Jeol). To determine the density of fibrillar packing of corneal collagen fibers, microphotographs were analyzed at magnification ×15,000. The number of collagen fibrils on an area of 250,000 nm² was calculated. The same microphotographs were used to measure the diameter of collagen fibrils. All numerical data were processed by the method of variational statistics. Histograms of the normal diameter distribution of collagen fibrils were analyzed. Results. The ultrastructure of corneal collagen on the 3rd day after UV corneal crosslinking was characterized by an increase in the interfibrillary space. This indicator was partially restored to its original values on the 90th day. At the same time, the fibrils retained the correct shape and sequence. The results of the analysis of the normal distribution of the diameter of collagen fibrils indicated an increase of an average of 10 %. Conclusion. An increase in the interfibrillary space after UV corneal crosslinking is probably associated with the development of postoperative stroma edema, which can be observed up to several months at the ultrastructural level. The revealed increase in the diameter of collagen fibrils is one of the factors increasing the strength and mechanical properties of the cornea. Keywords: UV cross-linking of rat cornea, electron microscopy, diameter of collagen fibrils, interfibrillary space.

Ultrastructural Characteristics of Chronically Failed Reconstructed Anterior Cruciate Ligament

Background In the present study, we have examined the ultrastructure of chronically failed reconstructed anterior cruciate ligament (ACL). We aimed to investigate a faulty ligamentization process of a failed reconstructed anterior cruciate ligament. In other words, we want to study ultrastructural alterations a failed ACL graft undergoes. Methods Two patients who underwent revision ACL reconstruction for nontraumatic failure without discontinuity of the graft were included in the study. The first patient was a 40-year-old male who had undergone ACL reconstruction of his right knee 21 years back using the bone-patellar tendon-bone autograft. The second patient was a 23-year-old male who had sustained an ACL tear with a medial collateral ligament injury treated by isolated ACL reconstruction 3 years back using hamstring tendon autograft. We collected punch biopsy specimens from the failed ligaments of both the patients during revision ACL reconstruction. These specimens were examined for the density of collagen fibrils within a fascicle (per 1.5 mm2), cellular metabolism, and fibril diameter (nm) by transmission electron microscopy. Results Fibroblasts of both the ligaments showed features of increased metabolism, more so in the first patient. Compared to the second patient, the fascicles of the first specimen were more loosely arranged. Both ligaments had a unimodal distribution of collagen fibrils. The first patient had a mean fibril diameter of 45.2 (+/−8.5) nm and an average fibril density of 376.8 fibrils per 1.5 mm2. The second patient had an average fibril diameter of 64.1 nm (+/−7) and a mean fibril density of 152.9 fibrils/1.5 mm2. The difference in these parameters of the two patients was statistically significant (P < 0.001). Conclusion Our study suggests that the absence of thicker collagen fibrils with unimodal distribution, the altered density of the collagen fibrils within a fascicle, and ovoid fibroblasts with increased metabolism may symbolize bad ligamentization changes.

Reticulocalbin 3 is involved in postnatal tendon development by regulating collagen fibrillogenesis and cellular maturation

Tendon plays a critical role in the joint movement by transmitting force from muscle to bone. This transmission of force is facilitated by its specialized structure, which consists of highly aligned extracellular matrix consisting predominantly of type I collagen. Tenocytes, fibroblast-like tendon cells residing between the parallel collagen fibers, regulate this specialized tendon matrix. Despite the importance of collagen structure and tenocyte function, the biological mechanisms regulating fibrillogenesis and tenocyte maturation are not well understood. Here we examine the function of Reticulocalbin 3 (Rcn3) in collagen fibrillogenesis and tenocyte maturation during postnatal tendon development using a genetic mouse model. Loss of Rcn3 in tendon caused decreased tendon thickness, abnormal tendon cell maturation, and decreased mechanical properties. Interestingly, Rcn3 deficient mice exhibited a smaller collagen fibril distribution and over-hydroxylation in C-telopeptide cross-linking lysine from α1(1) chain. Additionally, the proline 3-hydroxylation sites in type I collagen were also over-hydroxylated in Rcn3 deficient mice. Our data collectively suggest that Rcn3 is a pivotal regulator of collagen fibrillogenesis and tenocyte maturation during postnatal tendon development.

Safety and Long-term Scleral Biomechanical Stability of Rhesus Eyes after Scleral Cross-linking by Blue Light

ABSTRACT Purpose: To assess the safety and long-term scleral biomechanical stability of rhesus eyes after blue light scleral CXL by investigating the biomechanical and microstructural changes. Methods: Seven rhesus monkeys (14 eyes) were observed in this study. All right eyes received blue light scleral CXL at the superior temporal equatorial sclera, and the left eyes served as controls. Biological ocular parameters were followed up to 1 year after scleral CXL. Stress-strain measurements of three rhesus sclera were measured, three rhesus retinas were examined histologically by H&E and TUNEL staining. And the microstructure of both the sclera and retina were observed by transmission electron microscopy at 1 year. Results: As for the retinal thickness, choroidal thickness, flow density of retinal superficial vascular networks and flash electroretinography (f-ERG) results, no significant differences were observed between the paired eyes at 1 year (P >.05). At the same time, the scleral collagen fibril distribution was much tighter, and the scleral biomechanical properties were significantly increased in the experimental eyes. However, apoptotic cells and retinal ultrastructural changes could still be found in the retina of the experimental eyes. Conclusion: This study demonstrates that blue light scleral CXL could effectively increase the scleral stiffness of the rhesus eye for at least 1 year, but ultrastructural change was still observed in the retina of scleral CXL eye. Therefore, the long-term intraocular safety of the blue light scleral CXL technique for preventing myopia progression should be investigated further.

Stress–Strain Index Map: A New Way to Represent Corneal Material Stiffness

PurposeTo introduce a new method to map the mechanical stiffness of healthy and keratoconic corneas.MethodsNumerical modeling based on the finite element method was used to carry out inverse analysis of simulated healthy and keratoconic corneas to determine the regional variation of mechanical stiffness across the corneal surface based on established trends in collagen fibril distribution. The Stress–Strain Index (SSI), developed and validated in an earlier study and presented as a parameter that can estimate the overall stress–strain behavior of corneal tissue, was adopted in this research as a measure of corneal stiffness. The regional variation of SSI across the corneal surface was estimated using inverse analysis while referring to the common features of collagen fibrils’ distribution obtained from earlier x-ray scattering studies. Additionally, for keratoconic corneas, a method relating keratoconic cone features and cornea’s refractive power to the reduction in collagen fibril density inside the cone was implemented in the development of SSI maps. In addition to the simulated cases, the study also included two keratoconus cases, for which SSI maps were developed.ResultsSSI values varied slightly across corneal surface in the simulated healthy eyes. In contrast, both simulated and clinical keratoconic corneas demonstrated substantial reductions in SSI values inside the cone. These SSI reductions depended on the extent of the disease and increased with more considerable simulated losses in fibril density in the cone area. SSI values and their regional variation showed little change with changes in IOP, corneal thickness, and curvature.ConclusionSSI maps provide an estimation of the regional variation of biomechanical stiffness across the corneal surface. The maps could be particularly useful in keratoconic corneas, demonstrating the dependence of corneal biomechanical behavior on the tissue’s microstructure and offering a tool to fundamentally understand the mechanics of keratoconus progression in individual patients.

Fibril density reduction in keratoconic corneas.

This study aims to estimate the reduction in collagen fibril density within the central 6 mm radius of keratoconic corneas through the processing of microstructure and videokeratography data. Collagen fibril distribution maps and topography maps were obtained for seven keratoconic and six healthy corneas, and topographic features were assessed to detect and calculate the area of the cone in each keratoconic eye. The reduction in collagen fibril density within the cone area was estimated with reference to the same region in the characteristic collagen fibril maps of healthy corneas. Together with minimum thickness and mean central corneal refractive power, the cone area was correlated with the reduction in the cone collagen fibrils. For the corneas considered, the mean area of keratoconic cones was 3.30 ± 1.90 mm2. Compared with healthy corneas, fibril density in the cones of keratoconic corneas was lower by as much as 35%, and the mean reduction was 17 ± 10%. A linear approximation was developed to relate the magnitude of reduction to the refractive power, minimum corneal thickness and cone area (R2 = 0.95, p < 0.001). Outside the cone area, there was no significant difference between fibril arrangement in healthy and keratoconic corneas. The presented method can predict the mean fibril density in the keratoconic eye's cone area. The technique can be applied in microstructure-based finite-element models of the eye to regulate its stiffness level and the stiffness distribution within the areas affected by keratoconus.

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.

Effect of collagen fibril distributions on the crack profile in articular cartilage

Background and ObjectiveCartilage cracks and fissures may occur due to certain daily life activities such as sports practice, blunt trauma, and matrix fibrillation during early osteoarthritis. These cracks could further grow at the macroscopic level, alter the load distribution pattern in the matrix, limit the joint range of motion, and disturb chondrocytes synthesis. Cracks' shape and deformations in the loaded cartilage may affect the subsequent mechanobiological processes in the long term, likely because of the altered fluid exchange and excessive local deformations in the vicinity of the damage site. The fibrillar structure of the cartilage matrix appeared to have a protective effect against excessive deformations and tissue failure. Hence, in the present study, a fibril reinforced biphasic cartilage model was used to assess the potential role of different fibril orientations on the profile of a vertical crack in cartilage after applying a compressive load. MethodsA 20 × 20 × 1.5 mm3 cartilage model was developed with a 0.7 mm length V-shape cut at the center. Using an impermeable indenter, a 27% compression was applied to immature, mature, and isotropic cartilage models. Each of immature and mature groups had 4 different split line directions with respect to the cut edges, including 90°, 45°, 0°, and random orientation. The latter represented the disrupted collagen fibril orientations in early osteoarthritis. The model was verified with the experimental results in the literature. ResultsIn the superficial zones, the larger angle between the split lines and cut edges resulted in a wider cut opening. In the absence of collagen fibrils, the isotropic model resulted in a closed edge profile. Also, under a consistently applied compression, the OA model, with random collagen fibril distribution on its surface, had the smallest load-bearing capacity compared to the other models. ConclusionsFindings highlighted a primary role of collagen fibrils on the cut profile, which was more pronounced at dynamic rather than static conditions. Split lines perpendicular to the cut edges had some protective effects against the large dislocation of cut edges. These findings could be utilized to develop engineered tissues less susceptible to rupture. Moreover, the outcome of the present study can explain the potential causes of the crack propagation path reported in the literature.

Optical Microscopy and Electron Microscopy for the Morphological Evaluation of Tendons: A Mini Review.

The morphological characteristics of tendons have been thoroughly evaluated via microscopy. Optical microscopy and electron microscopy are the most commonly used techniques for tendon tissue observation. According to the principles of both microscopy types, preparation and evaluation methods vary. Simple optical microscopy is commonly used in the observation of cells and extracellular matrix, and many stains, including hematoxylin–eosin, Van Gieson, Prussian blue, Alcian blue, and toluidine blue, are used for evaluating cells, collagen fiber arrangement, and noncollagenous proteins. Histological scoring systems have been used in many studies for semi‐quantification. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) are the most commonly used electron microscopy types, and special consideration is needed for the fixation and embedding protocols. Glutaraldehyde followed by osmium is most commonly used in the chemical fixation of tendon tissue, followed by epoxy resin embedment. Longitudinal sections captured in SEM images show the arrangement of collagen fibrils and the cells and lipid drops among them, while cross sections captured in TEM images show the diameter and distribution of collagen fibrils. SEM and TEM are used together for comprehensive evaluations. This mini review is focused on the preparation methodology and related evaluation indexes for the morphological evaluation of tendons.

The pilocarpine-induced ciliary body contraction affects the elastic modulus and collagen of cornea and sclera in early development.

The aim of this study was to investigate the effects of pilocarpine-induced ciliary body constant contraction for a long time period on the elastic modulus and collagen in corneal and scleral tissues in the early developmental stage. Twelve one-month-old New Zealand white rabbits were randomly monocularly treated with pilocarpine to cause ciliary body constant contraction. After 1- and 2-months, the sclera tissues and the eyeballs were obtained to assess the cornea and three regions of the sclera-anterior, equatorial and posterior. The corneal tissues and the three regions of the scleral tissues were cut into strips for elastic modulus measurement using an Instron 5544, the size distribution of collagen fibrils was examined using electron microscopy, the samples were homogenized, and the concentration of hydroxyproline was measured to determine the collagen content. For corneal tissues, long-term pilocarpine-induced ciliary body constant contraction did not affect the elastic modulus and collagen. For scleral tissues, long-term pilocarpine-induced ciliary body constant contraction affected the elastic modulus, diameter of collagen fibrils and collagen content in the equatorial and posterior sclera, however, the anterior sclera were unaffected. These results suggested that pilocarpine-induced ciliary body contraction could affect the scleral structure and lead to deformation of the sclera and eyeball, thereby affecting visual functions function in the process of ocular emmetropization.

Histological properties of autogenous hamstring grafts after anterior cruciate ligament reconstruction

To investigate the histological characteristics of autogenous hamstring grafts after anterior cruciate ligament (ACL) reconstruction. The patients who underwent arthroscopic single-bundle ACL reconstruction with autogenous hamstring tendons and were followed up at least 4 years and also underwent second-look arthroscopy between March 2017 and December 2017 and met the selection criteria were considered for enrollment. Graft quality under arthroscopy was evaluated as good remodeling group (GRG, the total scores were 4-6) and poor remodeling group (PRG, the total scores were 1-3) according to synovial and vascular coverage, the apparent tension of the grafts, the thickness and retear of the grafts. During the second-look arthroscopic procedures, ACL graft biopsies were performed. Normal ACL tissues harvested from the patients under 60 years old who underwent total knee arthroplasty were designated as normal controls. Graft vascularity, cellular morphology, cellular metabolism, and collagen fibril distribution were analyzed. The 18 specimens (11 cases of GRG group and 7 cases of PRG group) and 9 native ACL biopsied tissue sample were enrolled into the study. Arthroscopy scores were 2-6 (mean, 4.7). The biology under light microscopy of GRG group was similar to that of native ACL in control group. There was no significant difference in the scores of graft vascularity and cellular morphology between GRG group and control group ( P>0.05), while PRG group was significantly lower than the other two groups ( P<0.05). Transmission electron microscope evaluation showed that GRG group and control group had better collagen fibril distribution and lower levels of cellular metabolism than PRG group ( P<0.05). There was no significant difference in cellular metabolism between GRG and control groups ( P>0.05), while collagen fibril distribution score of GRG group was significantly lower than that of control group ( P<0.05). While good remodeling grafts under arthroscopy in histological maturation period was proved to be more similar to normal ACL on ultrastructure properties under light and electron microscope, ultra structural differences regarding collagen fibril distribution still persist.