Collagen Fiber Bundles Research Articles

Fibrous finite element modeling of the optic nerve head region

The optic nerve head (ONH) region at the posterior pole of the eye is supported by a fibrous structure of collagen fiber bundles. Discerning how the fibrous structure determines the region biomechanics is crucial to understand normal physiology, and the roles of biomechanics on vision loss. The fiber bundles within the ONH structure exhibit complex three-dimensional (3D) organization and continuity across the various tissue components. Computational models of the ONH, however, usually represent collagen fibers in a homogenized fashion without accounting for their continuity across tissues, fibers interacting with each other and other fiber-specific effects in a fibrous structure. We present a fibrous finite element (FFE) model of the ONH that incorporates discrete collagen fiber bundles and their histology-based 3D organization to study ONH biomechanics as a fibrous structure. The FFE model was constructed using polarized light microscopy data of porcine ONH cryosections, representing individual fiber bundles in the sclera, dura and pia maters with beam elements and canal tissues as continuum structures. The FFE model mimics the histological in-plane orientation and width distributions of collagen bundles as well as their continuity across different tissues. Modeling the fiber bundles as linear materials, the FFE model predicts the nonlinear ONH response observed in an inflation experiment from the literature. The model also captures important microstructural mechanisms including fiber interactions and long-range strain transmission among bundles that have not been considered before. The FFE model presented here advances our understanding of the role of fibrous collagen structure in the ONH biomechanics. Statement of SignificanceThe microstructure and mechanics of the optic nerve head (ONH) are central to ocular physiology. Histologically, the ONH region exhibits a complex continuous fibrous structure of collagen bundles. Understanding the role of the fibrous collagen structure on ONH biomechanics requires high-fidelity computational models previously unavailable. We present a computational model of the ONH that incorporates histology-based fibrous collagen structure derived from polarized light microscopy images. The model predictions agree with experiments in the literature, and provide insight into important microstructural mechanisms of fibrous tissue biomechanics, such as long-range strain transmission along fiber bundles. Our model can be used to study the microstructural basis of biomechanical damage and the effects of collagen remodeling in glaucoma.

Structure, composition and anisotropic swelling of the bovine acetabular labrum

The fibro-cartilaginous labrum surrounds the acetabular rim and is important for hip joint stability and sealing. Sealing may be enhanced by swelling pressure within the normal labrum. Swelling of the degenerated or torn labrum might occur and potentially contribute to the development of osteoarthritis, through altered load transmission. This study aimed to characterize the three-dimensional swelling behaviour, the collagen fiber orientation and spatial proteoglycan distribution of the bovine acetabular labrum. Specimens were harvested from bovine donors (192–652 days, male, n = 6 donors). Structure was analyzed by scanning electron microscopy, histology, and dimethylmethylene blue assay. Specimen dimensions were measured before and after incubation in phosphate buffered saline to assess the swelling. Results showed that the articulating surface is composed of a collagen mesh network. Collagen fiber bundles showed a low degree of alignment close to the surface and were circumferentially aligned in the deep tissue. Proteoglycans were identified clustered between the collagen bundles. Glycosaminoglycan content was 10 x lower than that of cartilage (23.1 ± 6.4 compared to 299.5 ± 19.1 μg/mg dry weight) with minor regional differences. Specimens swelled significantly more in the orthogonal direction (swelling ratio 124.7 ± 10.2%) compared to the swelling parallel to the articulating surface (108.8 ± 6.1% and 102.8 ± 4.1%). In the deep tissue, swelling was also restricted in the main collagen fiber bundle direction (circumferentially), with a swelling ratio of 109.5 ± 4.0% in the main fiber bundle direction compared to 126.8 ± 7.3 % and 122.3 ± 5.8% radially. The findings demonstrate that the labrum shows anisotropic swelling properties, which reflect the anisotropy in the tissue structure and inter-fiber localisation of proteoglycans.

Direct measurements of collagen fiber recruitment in the posterior pole of the eye

Collagen is the main load-bearing component of the peripapillary sclera (PPS) and lamina cribrosa (LC) in the eye. Whilst it has been shown that uncrimping and recruitment of the PPS and LC collagen fibers underlies the macro-scale nonlinear stiffening of both tissues with increased intraocular pressure (IOP), the uncrimping and recruitment as a function of local stretch have not been directly measured. This knowledge is crucial to understanding their functions in bearing loads and maintaining tissue integrity. In this project we measured local stretch-induced collagen fiber bundle uncrimping and recruitment curves of the PPS and LC. Thin coronal samples of PPS and LC of sheep eyes were mounted and stretched biaxially quasi-statically using a custom system. At each step, we imaged the PPS and LC with instant polarized light microscopy and quantified pixel-level (1.5 μm/pixel) collagen fiber orientations. We used digital image correlation to measure the local stretch and quantified collagen crimp by the circular standard deviation of fiber orientations, or waviness. Local stretch-recruitment curves of PPS and LC approximated sigmoid functions. PPS recruited more fibers than the LC at the low levels of stretch. At 10% stretch the curves crossed with 75% bundles recruited. The PPS had higher uncrimping rate and waviness remaining after recruitment than the LC: 0.9º vs. 0.6º and 3.1º vs. 2.7º. Altogether our findings support describing fiber recruitment of both PPS and LC with sigmoid curves, with the PPS recruiting faster and at lower stretch than the LC, consistent with a stiffer tissue. Statement of significancePeripapillary sclera (PPS) and lamina cribrosa (LC) collagen recruitment behaviors are central to the nonlinear mechanical behavior of the posterior pole of the eye. How PPS and LC collagen fibers recruit under stretch is crucial to develop constitutive models of the tissues but remains unclear. We used image-based stretch testing to characterize PPS and LC collagen fiber bundle recruitment under local stretch. We found that fiber-level stretch-recruitment curves of PPS and LC approximated sigmoid functions. PPS recruited more fibers at a low stretch, but at 10% bundle stretch the two curves crossed with 75% bundles recruited. We also found that PPS and LC fibers had different uncrimping rates and non-zero waviness's when recruited.

Mice Lacking PLAP-1/Asporin Show Alteration of Periodontal Ligament Structures and Acceleration of Bone Loss in Periodontitis.

Periodontal ligament-associated protein 1 (PLAP-1), also known as Asporin, is an extracellular matrix protein expressed in the periodontal ligament and plays a crucial role in periodontal tissue homeostasis. Our previous research demonstrated that PLAP-1 may inhibit TLR2/4-mediated inflammatory responses, thereby exerting a protective function against periodontitis. However, the precise roles of PLAP-1 in the periodontal ligament (PDL) and its relationship to periodontitis have not been fully explored. In this study, we employed PLAP-1 knockout mice to investigate its roles and contributions to PDL tissue and function in a ligature-induced periodontitis model. Mandibular bone samples were collected from 10-week-old male C57BL/6 (WT) and PLAP-1 knockout (KO) mice. These samples were analyzed through micro-computed tomography (μCT) scanning, hematoxylin and eosin (HE) staining, picrosirius red staining, and fluorescence immunostaining using antibodies targeting extracellular matrix proteins. Additionally, the structure of the PDL collagen fibrils was examined using transmission electron microscopy (TEM). We also conducted tooth extraction and ligature-induced periodontitis models using both wild-type and PLAP-1 KO mice. PLAP-1 KO mice did not exhibit any changes in alveolar bone resorption up to the age of 10 weeks, but they did display an enlarged PDL space, as confirmed by μCT and histological analyses. Fluorescence immunostaining revealed increased expression of extracellular matrix proteins, including Col3, BGN, and DCN, in the PDL tissues of PLAP-1 KO mice. TEM analysis demonstrated an increase in collagen diameter within the PDL of PLAP-1 KO mice. In line with these findings, the maximum stress required for tooth extraction was significantly lower in PLAP-1 KO mice in the tooth extraction model compared to WT mice (13.89 N ± 1.34 and 16.51 N ± 1.31, respectively). In the ligature-induced periodontitis model, PLAP-1 knockout resulted in highly severe alveolar bone resorption, with a higher number of collagen fiber bundle tears and significantly more osteoclasts in the periodontium. Our results demonstrate that mice lacking PLAP-1/Asporin show alteration of periodontal ligament structures and acceleration of bone loss in periodontitis. This underscores the significant role of PLAP-1 in maintaining collagen fibrils in the PDL and suggests the potential of PLAP-1 as a therapeutic target for periodontal diseases.

Морфологическое обоснование применения альтернативного аллотрансплантата для офтальмохирургии

Purpose. Morphological assessment of fibrearchtechtonics of dispersed allografts made of cadaveric fascia lata. Material and methods. The morphological study of cadaveric fascia lata, made of its whole spirits-preserved and crushed grafts (10 samples each) was carried out. Light microscopy of collagen fibers (thickness of fiber bundles and gaps between them) in glycosaminoglycans (Hale's hystochemical reaction) was performed. Results. Studies of fascia lata samples which underwent technological processing showed that the tissue structure and its glycosaminoglycans are preserved; there was only a slight defibration of collagen fiber bundles. At the same time cellullar elements undergo complete destruction which reduces the immugenicity of allograft. It has been established that, in general, dispersed allografts have the same properties as native and preserved ones. Conclusion. Physicochemical processing of donor tissues, its crushing, fractioning and radiant sterilization allow to keep fibrearchtechtonics of fascia lata when making dispersed allografts, what is important for regenerative processes in ophthalmic surgery. Key words: morphological study, fibrearchtechtonics, fascia lata, structure, grafts, injectable biomaterials

Collagen Fibril Morphological Study of Medial Patellar Ligament of Stifle Joint Affected with Patellar Luxation in Cattle

Background: Upward fixation of patella is common clinical malady in cattle due to extensive movement of stifle joint during the locomotory activity because of unknown etiology. However, at certain peak point (overstretching of ligament) or due to occupational trauma (wear and tear), ligament become flaccid and weak leading to patellar displacement and lameness. Patellar ligaments play crucial role in stifle joint movement and any functional or structural alterations in the ligaments can lead to acute to chronic lameness in cattle. The present study was carried out to evaluate the structural changes in collagen fibril material in medial patellar ligament in cattle suffering with patellar luxation. Methods: Tissue specimens of medial patellar ligament of normal (control animal) and affected cattle (luxation of patella) were harvested and preserved for scanning electron microscopy (SEM) and transmission electron microscopic (TEM) study as per standard procedure at RUSKA Laboratories, College of Veterinary Science, PVNRTVU, Rajendranagar, Hyderabad, India. Result: The incidence of medial patellar luxation was higher in females as compared to male cattle and the duration of lameness was between 3-12 months (7.35±0.56 months) characterized by dragging of toe, outward jerky movement of affected limb, extension of limb and change in appearance of gait during walk and flexion of fetlock. Scanning electron study of normal ligament showed dense, uniformly and compact arrangement of collagen fiber bundles with morphometric thickness was 3.66±0.58 µm. whereas in luxated patella cases, medial ligament showed derangement in collagen fiber bundles. The collagen fibers were arranged irregular and wavy with morphometric thickness of collagen was 2.58±0.03 µm. Electron microscopy of the normal ligament showed presence of thick collagen fiber bundles packed in dense and compact arrangement. Electron microscopy of affected ligament had degenerative changes of the fibers characterized by thin collagen fiber bundles which were loosely and disorganized arranged fibrils. The present study concludes that, luxation of patella was common locomotory disorders in cattle due to derangement in collagen fiber bundles with drastic reduction in thickness of collagen fibers, increased gap between two collagen fibers, broken and overlapping fiber bundles and surface of collagen fibers appeared irregular rough via scanning and electron microscopy analysis.

Mesostructured Fibrils Exfoliated in Deep Eutectic Solvent as Building Blocks of Collagen Membranes.

The mesoscale components of collagen (nanofibrils, fibrils, and fiber bundles) are well organized in native tissues, resulting in superior properties and diverse functions. In this paper, we present a simple and controlled liquid exfoliation method to directly extract medium-sized collagen fibers ranging from 102 to 159 nm in diameter from bovine Achilles tendon using urea/hydrochloric acid and a deep eutectic solvent (DES). In situ observations under polarized light microscopy (POM) and molecular dynamics simulations revealed the effects of urea and GuHCl on tendon collagen. FTIR study results confirmed that these fibrils retained the typical structural characteristics of type I collagen. These shed collagen fibrils were then used as building blocks to create independent collagen membranes with good and stable mechanical properties, excellent barrier properties, and cell compatibility. A new method for collagen processing is provided in this work by using DES-assisted liquid exfoliation for constructing robust collagen membranes with mesoscale collagen fibrils as building blocks.

Construction of Human Periodontal Ligament Constitutive Model Based on Collagen Fiber Content.

Periodontal ligament (PDL) is mainly composed of collagen fiber bundles, and the content of collagen fiber is an important factor affecting the mechanical properties of PDL. Based on this, the purpose of this study is to explore the effect of the PDL collagen fiber content on its viscoelastic mechanical behavior. Transverse and longitudinal samples of different regions of PDL were obtained from the human maxilla. The fiber content at different regions of human PDL was quantitatively measured using image processing software, and a new viscoelastic constitutive model was constructed based on the fiber content. The nano-indentation experiment was carried out with a loading rate of 0.5 mN·s-1, a peak load of 3 mN, and a holding time of 200 s, and the model parameters were obtained through the experiment data. The results showed that with the increase of fiber content, the deformation resistance of PDL also increased, and compared with the neck and middle region, the compressive strain in the apical region of PDL was the largest. The range of reduced elastic modulus of human PDL was calculated to be 0.39~5.08 MPa. The results of the experimental data and the viscoelastic constitutive model fit well, indicating that the model can well describe the viscoelastic behavior of human PDL.

The effects of Shikonin on the hypertrophic scar of rabbit ears via the TLR4/NF-κB signaling pathway.

As a traditional Chinese medicine, Zihuang Shengji Ointment has obvious effects on promoting postoperative wound healing and reducing scar formation in clinical application. Shikonin is the major phytochemical in Zihuang Shengji Ointment. As a kind of naphquinone compound with anti-tumor, anti-viral, anti-inflammatory, anti-bacterial and other biological activities extracted from Lithospermum erythrorhizon, shikonin exerts an important role in many diseases. Shikonin has impacts on the development of hypertrophic scars (HS), however, these effects are yet mostly unknown. As a result, we created the Newland white rabbit ear HS model, administered shikonin to it, and then assessed scar hypertrophy using HE and VG staining. The degree of scarring is assessed by HI, NA, as well as AA. The expression levels of collagen I, collagen III, as well as α-SMA as well as fibroblast proliferation, are also measured using real-time PCR, immunohistochemistry, and western blot. TUNEL tests are used to assess fibroblast apoptosis. In our work, HE staining and VG staining showed that the shikonin-treated group had normal bundles of collagen fibers and regular fibroblasts. Shikonin suppresses the production of HS, according to histopathological features, HI, NA, and AA measures. Shikonin also causes fibroblast apoptosis and lowers the production of α-SMA, collagen I, as well as collagen III in the HS rat. Notably, we discover that NF-κB activation and TLR4 activity are inhibited by shikonin. Overall, the results show that the signaling pathway of TLR4/NF-κB is modulated by shikonin's inhibitory effect on scar formation, which represses the levels of collagen I, collagen III, α-SMA, as well as fibroblasts.

Effect of the Correction of Bilateral Differences in Masseter Muscle Functional Pressure on the Mandible of Growing Rats.

The objective of this study is to clarify the effect of restoring the lowered masticatory muscle functional pressure and correcting bilateral differences in masticatory muscle functional pressure on jawbone growth during growth and development with a quantitative evaluation of the changes in the micro/nanostructural characteristics of entheses. Male Wistar rats aged 4 weeks were divided into an experimental group injected with a botulinum toxin serotype A (BoNT/A) formulation to reduce muscle function (BTX group) and a control group (CTRL group). They were euthanised after 6, 8, 10, 12, and 16 weeks after measuring the difference between the midline of the upper and lower incisors. The mandibles were harvested for histological examination, second harmonic generation imaging, and the quantitative evaluation of biological apatite (BAp) crystal alignment. The midline difference decreased with age in weeks. In rats from 6 weeks after BoNT/A administration to 12 weeks after administration, the collagen fibre bundle diameter was significantly smaller in the BTX group; the difference between the two groups decreased with increasing age. BAp crystal alignment was significantly different on the x-axis and the y-axis on the BTX group from 6 weeks after BoNT/A administration to 10 weeks after administration. Asymmetry of mandibular bone formation caused by load imbalance during growth could be corrected by the adjustment of the function of the masseter muscle on either side.

The effects of orthodontic anchor screw inserted into the femur of growth-phase or mature rats -osteoid formation, bone mineral density, collagen fiber bundles, biological apatite crystal orientation.

The purpose of this study is to investigate the effect of orthodontic anchor screws (OASs) inserted into the femur of growth-phase or mature rats using histological observation and bone structure analysis. The experimental animals are growth-phase (6-week-old) or mature (25-week-old) male Wistar rats. OAS was placed into the point one-third of the femoral length from the proximal end of the femur, and the response of the surrounding bone was observed and measured. The results showed at the OAS bone interface, in growth-phase rats, bone mineral density (BMD) was reduced and the running angle of collagen fiber bundles varied significantly. In mature rats, more osteoid was observed and biological apatite (BAp) crystals showed a different orientation. It was suggested that after the insertion of OASs, bone volume and quality are decreased, but after a sufficient healing period, a new bone micro/nano structure, different from the original structure, are reconstructed.

Mammary myofibroblastoma: a clinicopathological analysis of fifteen cases

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of breast myofibroblastoma. Methods: The clinicopathological data and prognostic information of 15 patients with breast myofibroblastoma diagnosed at the Department of Pathology of the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China from 2014 to 2022 were collected. Their clinical characteristics, histological subtypes, immunophenotypes and molecular characteristics were analyzed. Results: There were 12 female and 3 male patients, ranging in age from 18 to 78 years, with a median and average age of 52 years. There were 6 cases in the left breast and 9 cases in the right breast, including 12 cases in outer upper quadrant, 2 cases in inner upper quadrant and 1 case in outer lower quadrant. Most of the cases showed a well-defined nodule grossly, including pushing growth under the microscope in 13 cases, being completely separated from the surrounding breast tissue in 1 case, and infiltrating growth in 1 case. Among them, 12 cases were classic subtype and composed of occasional spindle cells with varying intervals of collagen fiber bundles; eight cases had a small amount of fat; one case had focal cartilage differentiation; one case was epithelioid subtype, in which epithelioid tumor cells were scattered in single filing or small clusters; one case was schwannoma-like subtype, and the tumor cells were arranged in a significant palisade shape, resembling schwannoma, and one case was invasive leiomyoma-like subtype, in which the tumor cells had eosinophilic cytoplasm and were arranged in bundles, and infiltrating into the surrounding mammary lobules like leiomyoma. Immunohistochemical studies showed that the tumor cells expressed desmin (14/15) and CD34 (14/15), as well as ER (15/15) and PR (15/15). Three cases with histologic subtypes of epithelioid subtype, schwannoma-like subtype and infiltrating leiomyoma-like subtype showed RB1 negative immunohistochemistry. Then FISH was performed to detect RB1/13q14 gene deletion, and identified RB1 gene deletion in all three cases. Fifteen cases were followed up for 2-100 months, and no recurrence was noted. Conclusions: Myofibroblastoma is a rare benign mesenchymal tumor of the breast. In addition to the classic type, there are many histological variants, among which the epithelioid subtype is easily confused with invasive lobular carcinoma. The schwannoma-like subtype is similar to schwannoma, while the invasive subtype is easily misdiagnosed as fibromatosis-like or spindle cell metaplastic carcinoma. Therefore, it is important to recognize the various histological subtypes and clinicopathological features of the tumor for making correct pathological diagnosis and rational clinical treatment.

Alveolar Bone Microstructure Surrounding Orthodontic Anchor Screws with Plasma Surface Treatment in Rats.

A lateral load was applied to anchor screws that had undergone surface treatment, and the structure, cellular dynamics, and quality of the bone surrounding anchor screws were analyzed to investigate the effect of this surface treatment on the peri-implant jawbone. In addition, bone microstructural characteristics were quantitatively evaluated for each site of loading on the bone around the anchor screw. Rats were euthanized after observation on days 3, 5, or 7, and bone quality analyses were performed. Bone-implant contact rate increased more rapidly at an early stage in the treated surface group than in the untreated surface group. Bone lacuna morphometry showed that the measured values adjacent to the screw at the screw neck on the compressed side (A) and at the screw tip on the uncompressed side (D) were significantly lower than those at the screw tip on the compressed side (B) and at the screw neck on the uncompressed side (C). Collagen fiber bundle diameter showed that the measured values adjacent to regions A and D were significantly higher than those at regions B and C. Anchor screw surface activation facilitates initial bone contact of the screw, suggesting that early loading may be possible in clinical practice.

Morphofunctional characteristics of the skin in the assessment of adaptive capabilities in wild boars (sus scrofa scrofa (l., 1758)

The study of the mammalian skin reveals a wide range of adaptations of animals and their organs to various conditions of existence.The aim of the work is to establish, on the basis of factual material, the morphological and functional features of the skin of wild boars (Sus scrofa scrofa (Linnaeus 1758)) in conditions of adaptation to adverse natural and climatic factors.The skin samples of 6 adult individuals (males) of wild boar (Sus scrofa scrofa) caught in the autumn-winter period were studied. The morphometric parameters of the skin layers and their components were measured, based on the primary data, the total thickness of the skin and epidermis, the ratio of the outer and inner layers of the epidermis were calculated. The established absolute values of indicators (microns) were converted into relative values (%), the ratio of the stratum corneum and the underlying rows of the epidermis was found. The results obtained were processed using the Statistica (6.0) software package.In the studied animals, the skin is quite thick, with a wide variability of the indicator. We noted a well-developed epidermis in the skin of wild boars, its thickness averaged 72.99 microns. The stratum corneum, on average, reached a thickness of 36.13 microns (44.75-56.65% of total epidermis thickness), with a low level of variation (8.58%), which may be genetically deter mined. The dermis of the wild boars’ skin is well developed; it consists of densely packed, medium-sized bundles of collagen fibers that form ties with a heterogeneous binding, which indicates a high density and strength of the skin under mechanical stress and, possibly, is associated with sexual characteristics (the ability of the skin to withstand blows during fights) . Inclusions of adipose tissue were found in the lower layers of the dermis.The sebaceous glands are large, well developed, actively functioning in winter (as evidenced by the lipid mantle of the stratum corneum). A significant level of group variation was established, due to the wide individual variability of the indicators of functionally active structures (skin glands) in the studied animals, while the indicators of other derivatives of the epithelium – hair follicles, were distinguished by a very low level of variability.The data obtained allow us to conclude that the variability of the skin structure and its derivatives in wild boars have a certain significance in the process of adaptation to the effects of adverse climatic conditions, depending on the season of the year.

Multifilament Collagen Fiber Bundles with Tendon-like Structure and Mechanical Performance.

Collagen multifilament bundles comprised of thousands of monofilaments are prepared by multipin contact drawing of an entangled polymer solution consisting of collagen and poly(ethylene oxide) (PEO). The multifilament bundles are hydrated in graded concentrations of PEO and phosphate buffered saline (PBS) to promote assembly of collagen fibrils within each monofilament while preserving the structure of the multifilament bundle. Multiscale structural characterization reveals that the hydrated multifilament bundle contains properly folded collagen molecules packed in collagen fibrils containing microfibrils, staggered by exactly one-sixth of the microfibril D-band spacing to produce a periodicity of 11nm. Sequence analysis predicts that in this structure, phenylalanine residues are close enough within and between microfibrils to become ultraviolet C (UVC) crosslinked. In agreement with this analysis, the ultimate tensile strength (UTS) and Young's modulus of the hydrated collagen multifilament bundles crosslinked by UVC radiation increase nonlinearly with total UVC energy to reach values in the range of native tendons without damage to the collagen molecules. This fabrication method recapitulates the structure of a tendon across multiple length scales and offers tunability in tensile properties using only collagen molecules and no other chemical additives in addition to PEO, which is almost entirely removed during the hydration process.

MICROSCOPIC CHANGES IN RAT’S GINGIVA UNDER CONDITIONS OF EXPERIMENTAL PERIODONTITIS AND THE USE OF MESENCHYMAL STEM CELLS

MICROSCOPIC CHANGES IN RAT’S GINGIVA UNDER CONDITIONS OF EXPERIMENTAL PERIODONTITIS AND THE USE OF MESENCHYMAL STEM CELLS

Immunohistochemical analysis of the ligamentum mucosum is the key to understand its clinical usefulness

Ligamentum Mucosum(LM) is a ligamentous structure within the synovial layer of the knee joint capsule. For a long time LM was perceived as the vestigial remnant from the embryological development of the knee. Being treated as an irrelevant structure LM was often the first victim of shaver during arthroscopy. However, last years have shown increased interest in this structure due to its potentially significant clinical role. Our aim was to classify LM based on its morphological characteristics and examine its microanatomy usingimmunohistochemical analysis to reveal the potential clinical value for surgeons. We have examined sixteen fresh frozen lower limbs, 6 females (mean age 83.1 ± 3.4 years) and 10 males (mean age 84.2 ± 6.8 years). Classical histological H+E stain was routinely conducted. Subsequently, CD31 antibody (DAKO, Monoclonal Mouse Anti-Human, Clone JC70A) was used to mark vascular epithelium. Monoclonal Mouse Anti-Human Neurofilament Protein (NFP) antibody (DAKO, Clone 2F11) was used to expose the nerves. Moreover, we have conducted arthroscopic visualizing and suturing LM to the torn ACL during routinely performed arthroscopic suturing of the ACL. The dissection process has revealed that LM was present only in 75% of cases.Histological examination confirmed the presence of longitudinal collagen fiber bundles in all samples. Tiny nerves were confirmed by NFP, along the subsynovial layer in all samples. CD-31 immunostain revealed the presence of many vascular vessels along the entire ligament, especially well developed at its distal end. Our study has revealed that LM contains rich vascular network. Thus, it may be a donor for the revascularization process after ACL tear or reconstruction which may improve the recovery. Another great advantage of the LM is the presence of nerves along the subsynovial layer, hopefully they may serve as the source of reinnervation and hence better clinical outcome. Based on our results we believe that seemingly irrelevant LM may be very useful during surgical procedures in the knee region. Suturing LM to the ACL may not only prevent the infrapatellar fat pad from subluxation but also improve the blood flow and reinnervation of the injured ACL. Until now there are only a few studies examining microanatomy of the LM. This basic knowledge may serve as the foundation for surgical procedures. Hopefully our findings may be useful for surgeons while planning surgical procedures or clinicians while diagnosing patients who suffer anterior knee pain.

Ultraviolet light induces mechanical and structural changes in full thickness human skin

While the detrimental health effects of prolonged ultraviolet (UV) irradiation on skin health have been widely accepted, the biomechanical process by which photoaging occurs and the relative effects of irradiation with different UV ranges on skin biomechanics have remained relatively unexplored. In this study, the effects of UV-induced photoageing are explored by quantifying the changes in the mechanical properties of full-thickness human skin irradiated with UVA and UVB light for incident dosages up to 1600 J/cm2. Mechanical testing of skin samples excised parallel and perpendicular to the predominant collagen fiber orientation show a rise in the fractional relative difference of elastic modulus, fracture stress, and toughness with increased UV irradiation. These changes become significant with UVA incident dosages of 1200 J/cm2 for samples excised both parallel and perpendicular to the dominant collagen fiber orientation. However, while mechanical changes occur in samples aligned with the collagen orientation at UVB dosages of 1200 J/cm2, statistical differences in samples perpendicular to the collagen orientation emerge only for UVB dosages of 1600 J/cm2. No notable or consistent trend is observed for the fracture strain. Analyses of toughness changes with maximum absorbed dosage reveals that no one UV range is more impactful in inducing mechanical property changes, but rather these changes scale with maximum absorbed energy. Evaluation of the structural characteristics of collagen further reveals an increase in collagen fiber bundle density with UV irradiation, but not collagen tortuosity, potentially linking mechanical changes to altered microstructure.

Analysis of monocyte subpopulations with cardiovascular, burn and other pathologies (2010 classification)

A classification of monocyte subpopulations developed in 2010 with coverage of the immunobiological properties of cells, their functional activity and participation in various pathological processes (inflammatory, cardiovascular diseases, strokes, myocardial infarctions, aortic aneurysms, surgical modification of heart valves, diabetes, burns, etc.). The diagnostic and prognostic aspects of the analysis of monocytic subpopulations are considered. The unique data obtained by the staff of the Gamaleya Institute of epidemiology and microbiology. They consist in the fact that various physical forms of unmodified native type 1 collagen are powder, i.e. crushed bundles of collagen fibers, a hydrogel or a solution of extracellular matrix peptides, as well as a suspension of collagen fibers obtained from a powder, when applied to the surface of acute and chronic and diabetic wounds, bedsores, trophic ulcers, etc., can provide a pronounced anti-inflammatory, reparative, remodulating and regenerative effect on condition of wounds, providing their accelerated healing due to the local accumulation of “regenerative” subpopulations of Mon3 monocytes, which can be most directly used in burn tissue lesions. In this case, the analysis of monocytic subpopulations is of paramount importance. Moreover, possible potentiating effects of additional use in burns under the control of the analysis of monocyte subpopulations of powerful modern pluripotent immunomodulators – polyoxidonium, galavit and their possible combination with local use of collagen preparations are discussed. Finally, we obtained preliminary data indicating the development in burned patients of a deficiency in the absolute and relative content of the most important “patrolling” non-classical subpopulation of CD14+CD16++ monocytes compared with healthy primary (non-professional) donors, which can be a very important finding in the diagnosis and prognosis and substantiation of new methods of treatment of burns.

Changes in the microstructure of the human aortic adventitia under biaxial loading investigated by multi-photon microscopy.

Among the three layers of the aortic wall, the media is primarily responsible for its mechanical properties, but the adventitia prevents the aorta from overstretching and rupturing. The role of the adventitia is therefore crucial with regard to aortic wall failure, and understanding the load-induced changes in tissue microstructure is of high importance. Specifically, the focus of this study is on the changes in collagen and elastin microstructure in response to macroscopic equibiaxial loading applied to the aortic adventitia. To observe these changes, multi-photon microscopy imaging and biaxial extension tests were performed simultaneously. In particular, microscopy images were recorded at 0.02 stretch intervals. The microstructural changes of collagen fiber bundles and elastin fibers were quantified with the parameters of orientation, dispersion, diameter, and waviness. The results showed that the adventitial collagen was divided from one into two fiber families under equibiaxial loading conditions. The almost diagonal orientation of the adventitial collagen fiber bundles remained unchanged, but the dispersion was substantially reduced. No clear orientation of the adventitial elastin fibers was observed at any stretch level. The waviness of the adventitial collagen fiber bundles decreased under stretch, but the adventitial elastin fibers showed no change. These original findings highlight differences between the medial and adventitial layers and provide insight into the stretching process of the aortic wall. STATEMENT OF SIGNIFICANCE: To provide accurate and reliable material models, it is essential to understand the mechanical behavior of the material and its microstructure. Such understanding can be enhanced with tracking of the microstructural changes caused by mechanical loading of the tissue. This study provides therefore a unique dataset of structural parameters of the human aortic adventitia obtained under equibiaxial loading. The structural parameters describe orientation, dispersion, diameter, and waviness of collagen fiber bundles and elastin fibers. Eventually, the microstructural changes in the human aortic adventitia are compared with the microstructural changes in the human aortic media from a previous study. This comparison reveals the cutting-edge findings on the differences in the response to the loading between these two human aortic layers.