Ultrasound pathology of the entheses in an age and gender stratified sample of healthy adult subjects: a prospective cross-sectional frequency study.

OBJECTIVES: Ultrasound (US) examination of the entheses is increasingly used. However, little is known about US findings in the entheses in asymptomatic persons. The aim of this study was to investigate the appearance of US signs in the enthuses of the lower limb in asymptomatic subjects. METHODS: We recruited 64 subjects, eight women and eight men whose ages covered four decades, from 20 to 60 years. None had tendon or joint disease in the lower limbs. Participants were examined by a rheumatologist and blood samples were collected to rule out enthesis pathology. The enthesis of the dominant leg were examined with grey-scale and Doppler US to evaluate increased thickness, changed structure, enthesophytes/calcifications, erosions, and colour Doppler signal. RESULTS: Ultrasound examination of 320 entheses was made. At enthesis level, elementary lesions were seen at 73 (22.8%) sites, at subject-level 47 (73.4%) persons showed elementary lesions, in 27 (57%) only one enthesis was affected. Doppler activity was seen in four sites, three at the quadriceps insertion. Most common US elementary lesion was enthesophytes at the Achilles and quadriceps tendon insertion. A tendency towards more elementary lesions was seen in men, and a slight increase was seen with increasing age, however, not statistically significance. CONCLUSIONS: Our findings suggest that US can be used to diagnose/examine subjects in adulthood for pathological changes in the entheses; however, caution should be taken regarding enthesophytes of the quadriceps and Achilles tendon.

Development of fibrocartilage layers in the anterior cruciate ligament insertion in rabbits.

BACKGROUND: A detailed evaluation focusing on the fibrocartilage layers in the anterior cruciate ligament (ACL) insertion is necessary to consider regeneration of the insertion. This study examined the development of the fibrocartilage layers in the ACL tibial insertion in rabbits by quantitative morphometric evaluations based on histological and immunohistochemical analyses. METHODS: Male Japanese white rabbits were used because of their history of use for histomorphometric analyses of the ACL insertion and to eliminate the influence of female hormones on the ACL. Six animals were euthanized at each age (1 day and 1, 2, 4, 6, 8, 12, and 24 weeks); in total, 48 animals were used. Proliferation rate, apoptosis rate, Sox9-positive rate, and chondrocyte number were evaluated. Safranin O-stained glycosaminoglycan (GAG) areas, tidemark length, ACL insertion width, and ACL length were also evaluated. All parameters were compared with those at age 24 weeks of age. RESULTS: High levels of chondrocyte proliferation and Sox9 expression continued until 4 and 8 weeks of age, respectively, and then gradually decreased. Chondrocyte apoptosis increased up to 8 weeks. The chondrocyte number, ACL insertion width, ACL length, safranin O-stained GAG areas, and tidemark length gradually increased up to 12 weeks. CONCLUSION: Chondrocytes that displayed chondrocyte proliferation and Sox9 expression increased until 12 weeks of age, in accordance with development of the ACL length and its insertion width. The GAG production and tidemark length also increased until 12 weeks of age. The development of fibrocartilage layers in the ACL insertion was complete at 12 weeks of age.

A more flattened bone tunnel has a positive effect on tendon-bone healing in the early period after ACL reconstruction.

PURPOSE: The purpose of this study was to evaluate whether a flattened bone tunnel has a positive effect on the tendon-bone healing (TBH) process in the early period after anterior cruciate ligament (ACL) reconstruction. METHODS: Seventy-two New Zealand White rabbits were randomly allocated into two groups, the flattened tunnel (FT) group and the conventional round tunnel (RT) group. We compared the cross-sectional areas and diameters of the bone tunnels between the two groups through computed tomography (CT) scanning. TBH results between the two groups were assessed by histological analysis, micro-CT scanning and biomechanical tests at 4 weeks, 8 weeks and 12 weeks after operation. RESULTS: The cross-sectional areas of the bone tunnels between the two groups were almost the same. However, the shape of bone tunnels in the FT group was more flattened. A faster cellular and collagen remoulding process were found in the FT group. Semiquantitative histological analysis of Safranin O staining showed that there was more fibrocartilage formation in the interface region in the FT group (P < 0.05). Sirius Red staining showed that the tissues in the interface areas were more intense in the FT group. Micro-CT scanning showed that more new bone formation could be found in the interface region in the FT group. The biomechanical tests also showed that FT ACL reconstruction will result in a stronger regenerated tendon-bone interface. CONCLUSIONS: Our study found that a flattened bone tunnel accelerated TBH in the early period after ACL reconstruction surgery in a rabbit model, which lays the groundwork for further clinical practice of this ACL reconstruction method.

Human Tendon-Derived Collagen Hydrogel Significantly Improves Biomechanical Properties of the Tendon-Bone Interface in a Chronic Rotator Cuff Injury Model.

PURPOSE: Poor healing of the tendon-bone interface (TBI) after rotator cuff (RTC) tears leads to high rates of recurrent tear following repair. Previously, we demonstrated that an injectable, thermoresponsive, type I collagen-rich, decellularized human tendon-derived hydrogel (tHG) improved healing in an acute rat Achilles tendon injury model. The purpose of this study was to investigate whether tHG enhances the biomechanical properties of the regenerated TBI in a rat model of chronic RTC injury and repair. METHODS: Tendon hydrogel was prepared from chemically decellularized human cadaveric flexor tendons. Eight weeks after bilateral resection of supraspinatus tendons, repair of both shoulders was performed. One shoulder was treated with a transosseous suture (control group) and the other was treated with a transosseous suture plus tHG injection at the repair site (tHG group). Eight weeks after repair, the TBIs were evaluated biomechanically, histologically, and via micro-computed tomography (CT). RESULTS: Biomechanical testing revealed a larger load to failure, higher stiffness, higher energy to failure, larger strain at failure, and higher toughness in the tHG group versus control. The area of new cartilage formation was significantly larger in the tHG group. Micro-CT revealed no significant difference between groups in bone morphometry at the supraspinatus tendon insertion, although the tHG group was superior to the control. CONCLUSIONS: Injection of tHG at the RTC repair site enhanced biomechanical properties and increased fibrocartilage formation at the TBI in a chronic injury model. CLINICAL RELEVANCE: Treatment of chronic RTC injuries with tHG at the time of surgical treatment may improve outcomes after surgical repair.

Part I: Development and Physiology of the Temporomandibular Joint.

PURPOSE OF REVIEW: Investigate the developmental physiology of the temporomandibular joint (TMJ), a unique articulation between the cranium and the mandible. RECENT FINDINGS: Principal regulatory factors for TMJ and disc development are Indian hedgehog (IHH) and bone morphogenetic protein (BMP-2). The mechanism is closely associated with ear morphogenesis. Secondary condylar cartilage emerges as a subperiosteal blastema on the medial surface of the posterior mandible. The condylar articular surface is immunoreactive for tenascin-C, so it is a modified fibrous periosteum with an underlying proliferative zone (cambrium layer) that differentiates into fibrocartilage. The latter cushions high loads and subsequently produces endochondral bone. The TMJ is a heavily loaded joint with three cushioning layers of fibrocartilage in the disc, as well as in subarticular zones in the fossa and mandibular condyle. The periosteal articular surface produces fibrocartilage to resist heavy loads, and has unique healing and adaptive properties for maintaining life support functions under adverse environmental conditions.

Bridging Repair of Large Rotator Cuff Tears Using a Multilayer Decellularized Tendon Slices Graft in a Rabbit Model.

PURPOSE: The purpose of this study was to evaluate the efficacy of an extracellular matrix scaffold with multilayer decellularized tendon slices (MDTSs) for reconstructing large rotator cuff tears in a rabbit model. METHODS: Large defects in the infraspinatus tendons were created bilaterally in 36 rabbits. The graft group underwent bridging repair of the defects with the MDTSs grafts from Achilles tendons of adult beagle dogs, and the control group underwent repair with the autologous excised tendon. Specimens underwent histologic observation, biomechanical testing, and microcomputed tomography analysis at 2, 4, and 8 weeks after surgery. RESULTS: Histologic analysis confirmed that the MDTSs graft promoted cell ingrowth and tissue integration, and fibrocartilage and Sharpey fibers formed at the enthesis at 8 weeks. Accordingly, the MDTSs graft generated a histologic appearance similar to that of the autogenous tendon graft. Mechanical testing revealed a significant increase of the regenerated tendons in ultimate load and stiffness from 4 to 8 weeks postoperatively, which was similar to autologous tendon repair. Microcomputed tomography analysis demonstrated that the MDTSs graft promoted bone formation at the tendon-bone insertion, thus improving the mechanical properties of the repair tendon. CONCLUSIONS: The MDTSs graft used to bridge large rotator cuff defects in a rabbit model promoted host cell ingrowth, enhanced the remodeling of regenerated tendon, and promoted fibrocartilage formation, thus improving the biomechanical properties of the repaired tendon. This study thereby provides fundamental information for rotator cuff regeneration with the MDTSs graft. CLINICAL RELEVANCE: Rotator cuff regeneration using MDTSs grafts is a promising procedure for large rotator cuff tears.

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

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

Biomechanical evaluation contribution of the acetabular labrum to hip stability.

PURPOSE: Knowledge of the effect of hip pathologies on hip biomechanics is important to the understanding of the development of osteoarthritis, and the contribution of the labrum to hip joint stability has had limited study. The purpose of this study was to evaluate the effect of labral injury to stability of the femoral head in the acetabular socket. METHODS: Ten cadaver hip specimens were tested using a robotic system under four different loading conditions: axial loading (80 N) along the femoral axis and axial loading (80 N) combined with either anterior, posterior or lateral loading (60 N). The hip states were examined were intact, with a 1.5 cm capsulotomy and with a 1 cm resection of the anterosuperior labrum. RESULTS: At 30° of flexion, under axial load, the displacement of the hip with capsulotomy and labral resection (9.6 ± 2.5 mm) was significantly larger then the hip with capsulotomy alone (5.6 ± 4.1 mm, p = 0.005) and the intact hip (5.2 ± 3.8 mm, p = 0.005). Also, at 30° of flexion, the displacement under combined axial and anterior/posterior load was increased with capsulotomy and labral resection. CONCLUSION: The acetabular labrum provides stability to the hip joint in response to a distraction force and combined distraction and translation forces. One centimetre of labral resection caused significant displacement ("wobbling" effect) of the femoral head within the acetabulum with normal range of motion. Successful labral repair could be crucial for restoration of the hip biomechanics and prevention of coxarthrosis.

Immortalization and characterization of mouse temporomandibular joint disc cell clones with capacity for multi-lineage differentiation.

OBJECTIVE: Despite the importance of temporomandibular joint (TMJ) disc in normal function and disease, studying the responses of its cells has been complicated by the lack of adequate characterization of the cell subtypes. The purpose of our investigation was to immortalize, clone, characterize and determine the multi-lineage potential of mouse TMJ disc cells. DESIGN: Cells from 12-week-old female mice were cultured and immortalized by stable transfection with human telomerase reverse transcriptase (hTERT). The immortalized cell clones were phenotyped for fibroblast- or chondrocyte-like characteristics and ability to undergo adipocytic, osteoblastic and chondrocytic differentiation. RESULTS: Of 36 isolated clones, four demonstrated successful immortalization and maintenance of stable protein expression for up to 50 passages. Two clones each were initially characterized as fibroblast-like and chondrocyte-like on the basis of cell morphology and growth rate. Further the chondrocyte-like clones had higher mRNA expression levels of cartilage oligomeric matrix protein (COMP) (>3.5-fold), collagen X (>11-fold), collagen II expression (2-fold) and collagen II:I ratio than the fibroblast-like clones. In contrast, the fibroblast-like clones had higher mRNA expression level of vimentin (>1.5-fold), and fibroblastic specific protein 1 (>2.5-fold) than the chondrocyte-like clones. Both cell types retained multi-lineage potential as demonstrated by their capacity to undergo robust adipogenic, osteogenic and chondrogenic differentiation. CONCLUSIONS: These studies are the first to immortalize TMJ disc cells and characterize chondrocyte-like and fibroblast-like clones with retained multi-differentiation potential that would be a valuable resource in studies to dissect the behavior of specific cell types in health and disease and for tissue engineering.

Sacrificial nanofibrous composites provide instruction without impediment and enable functional tissue formation.

The fibrous tissues prevalent throughout the body possess an ordered structure that underlies their refined and robust mechanical properties. Engineered replacements will require recapitulation of this exquisite architecture in three dimensions. Aligned nanofibrous scaffolds can dictate cell and matrix organization; however, their widespread application has been hindered by poor cell infiltration due to the tight packing of fibers during fabrication. Here, we develop and validate an enabling technology in which tunable composite nanofibrous scaffolds are produced to provide instruction without impediment. Composites were formed containing two distinct fiber fractions: slow-degrading poly(ε-caprolactone) and water-soluble, sacrificial poly(ethylene oxide), which can be selectively removed to increase pore size. Increasing the initial fraction of sacrificial poly(ethylene oxide) fibers enhanced cell infiltration and improved matrix distribution. Despite the removal of >50% of the initial fibers, the remaining scaffold provided sufficient instruction to align cells and direct the formation of a highly organized ECM across multiple length scales, which in turn led to pronounced increases in the tensile properties of the engineered constructs (nearly matching native tissue). This approach transforms what is an interesting surface phenomenon (cells on top of nanofibrous mats) into a method by which functional, 3D tissues (>1 mm thick) can be formed, both in vitro and in vivo. As such, this work represents a marked advance in the engineering of load-bearing fibrous tissues, and will find widespread applications in regenerative medicine.

Ultrasound of the joints and entheses in healthy children.

BACKGROUND: Tendon insertion pathologies such as enthesitis and apophysitis in children can result from trauma, overuse syndrome and arthritis. Knowledge of the US appearance of normal joints by age might aid diagnosis of pathologies. OBJECTIVE: We describe the age-related sonographic features of the elbows, knees and feet in healthy children, providing a reference for the normal appearance of tendon insertions, apophyseal cartilage and bursae. MATERIALS AND METHODS: This is a prospective cross-sectional study of 30 healthy children. Children were grouped according to age: group 1 (4-9 years, n = 11), group 2 (10-13 years, n = 9) and group 3 (14-18 years, n = 10). Children completed pain and function questionnaires and underwent a standardized joint examination by a pediatric rheumatologist. The common extensor, common flexor, quadriceps, patellar and Achilles tendons and plantar fascia insertions were evaluated with gray-scale and power Doppler ultrasound. The anterior elbow, suprapatellar and retrocalcaneal bursae were evaluated for fluid. We measured the apophyseal cartilage thickness at the enthesis. Correlation analyses examined associations between age and tendon thickness. We used ANOVA, with location as a repeated measure, to test for gender differences in cartilage thickness. RESULTS: Children had a median age of 12.4 years and 55% were boys. All 360 entheses appeared normal on gray-scale imaging. There was a strong linear relationship between tendon thickness and age. Tendon vascularity was only present in young children (group 1), in 7/22 (32%) quadriceps tendons. Peri-tendinous power Doppler signal was seen at seven sites: two patellar, four quadriceps and one common flexor tendon, and all these children were in group 2. Suprapatellar bursal fluid <3 mm was detected in 9/60 (15%) knees. Of the children in group 1, boys had thicker apophyseal cartilage than girls at the medial epicondyle, patellar poles and os calcis (P < 0.05). CONCLUSION: Tendon vascularity may be a normal finding in young children, and mild peri-tendinous vascularity is not uncommon in children 10-13 years of age. Tendon thickness has a linear relationship with age; however cartilage thickness varies across sites and also differs as a function of gender.

Engineering anisotropic biomimetic fibrocartilage microenvironment by bioprinting mesenchymal stem cells in nanoliter gel droplets.

Over the past decade, bioprinting has emerged as a promising patterning strategy to organize cells and extracellular components both in two and three dimensions (2D and 3D) to engineer functional tissue mimicking constructs. So far, tissue printing has neither been used for 3D patterning of mesenchymal stem cells (MSCs) in multiphase growth factor embedded 3D hydrogels nor been investigated phenotypically in terms of simultaneous differentiation into different cell types within the same micropatterned 3D tissue constructs. Accordingly, we demonstrated a biochemical gradient by bioprinting nanoliter droplets encapsulating human MSCs, bone morphogenetic protein 2 (BMP-2), and transforming growth factor ß1 (TGF- ß1), engineering an anisotropic biomimetic fibrocartilage microenvironment. Assessment of the model tissue construct displayed multiphasic anisotropy of the incorporated biochemical factors after patterning. Quantitative real time polymerase chain reaction (qRT-PCR) results suggested genomic expression patterns leading to simultaneous differentiation of MSC populations into osteogenic and chondrogenic phenotype within the multiphasic construct, evidenced by upregulation of osteogenesis and condrogenesis related genes during in vitro culture. Comprehensive phenotypic network and pathway analysis results, which were based on genomic expression data, indicated activation of differentiation related mechanisms, via signaling pathways, including TGF, BMP, and vascular endothelial growth factor.

The role of extracellular matrix elasticity and composition in regulating the nucleus pulposus cell phenotype in the intervertebral disc: a narrative review.

Intervertebral disc (IVD) disorders are a major contributor to disability and societal health care costs. Nucleus pulposus (NP) cells of the IVD exhibit changes in both phenotype and morphology with aging-related IVD degeneration that may impact the onset and progression of IVD pathology. Studies have demonstrated that immature NP cell interactions with their extracellular matrix (ECM) may be key regulators of cellular phenotype, metabolism and morphology. The objective of this article is to review our recent experience with studies of NP cell-ECM interactions that reveal how ECM cues can be manipulated to promote an immature NP cell phenotype and morphology. Findings demonstrate the importance of a soft (<700 Pa), laminin-containing ECM in regulating healthy, immature NP cells. Knowledge of NP cell-ECM interactions can be used for development of tissue engineering or cell delivery strategies to treat IVD-related disorders.

Viability assessment of the chondral flap in patients with cam-type femoroacetabular impingement: a preliminary report.

BACKGROUND: Delaminated acetabular cartilage is a common finding in patients undergoing surgical dislocation or hip arthroscopy in the treatment of cam-type femoroacetabular impingement. Current treatment involves resection of the free cartilage flap with or without acetabular rim trimming. The viability of the delaminated cartilage flap is not known. We sought to examine if the acetabular cartilage still has viable cartilage cells and, if so, what type of cartilage is present. METHODS: We examined the delaminated cartilage flaps from patients undergoing surgical dislocation and osteochondroplasty for symptomatic cam-type impingement. We performed hematoxylin and eosin staining and histological analysis using light microscopy to determine cartilage viability and cartilage type. RESULTS: We examined 12 delaminated cartilage flaps from 11 patients (10 men, 1 woman, average age 30.1 yr). Ninety percent chondrocyte viability was confirmed in 11 of 12 flaps. Six of 12 flaps were composed predominantly of hyaline cartilage, 4 were a mixed population of fibrocartilage and hyaline cartilage and 2 were predominantly fibrocartilage. CONCLUSION: Our findings suggest that the delaminated cartilage flap in patients with femoroacetabular impingement may retain a large amount of viable chondrocytes. Development of surgical techniques focusing on refixation of this flap as an alternative to excision and microfracture should be considered.

CONTEXTE: La présence d'un cartilage acétabulaire délaminé s'observe souvent chez les patients qui subissent une dislocation chirurgicale ou une arthroscopie de la hanche pour le traitement du conflit fémoro-acétabulaire de type came. Le traitement actuel repose sur la résection du lambeau articulaire libre, avec ou sans résection du rebord acétabulaire. La viabilité du lambeau de cartilage délaminé est inconnue. Nous avons voulu vérifier si le cartilage acétabulaire conserve des cellules de cartilage viables et le cas échéant, quel type de cartilage est présent. MÉTHODES: Nous avons examiné les lambeaux de cartilage délaminés provenant de patients soumis à une dislocation et ostéochondroplastie chirurgicales pour un conflit de type came symptomatique. Nous avons procédé à une coloration à l'hématoxyiline et à l'éosine, ainsi qu'à une analyse histologique par microscopie optique pour déterminer le type de cartilage et sa viabilité. RÉSULTATS: Nous avons examiné 12 lambeaux de cartilage délaminé provenant de 11 patients (10 hommes, 1 femme, âgés en moyenne de 30,1 ans). La viabilité des chondrocytes a été confirmée à 90 % pour 11 lambeaux sur 12. Six lambeaux sur 12 se composaient surtout de cartilage hyalin, 4 étaient un mélange de fibrocartilage et de cartilage hyalin et 2 étaient principalement du fibrocartilage. CONCLUSION: Selon nos observations, le lambeau de cartilage délaminé chez les patients qui présentent un conflit fémoro-acétabulaire peut conserver une forte proportion de chondrocytes viables. Il faut envisager la mise au point de techniques chirurgicales axées sur la « refixation ¼ de ce lambeau comme solution de rechange à l'exérèse et à la microfracture.

Compositional, Structural, and Biomechanical Properties of Three Different Soft Tissue-Hard Tissue Insertions: A Comparative Review.

Connective tissue attaches to bone across an insertion with spatial gradients in components, microstructure, and biomechanics. Due to regional stress concentrations between two mechanically dissimilar materials, the insertion is vulnerable to mechanical damage during joint movements and difficult to repair completely, which remains a significant clinical challenge. Despite interface stress concentrations, the native insertion physiologically functions as the effective load-transfer device between soft tissue and bone. This review summarizes tendon, ligament, and meniscus insertions cross-sectionally, which is novel in this field. Herein, the similarities and differences between the three kinds of insertions in terms of components, microstructure, and biomechanics are compared in great detail. This review begins with describing the basic components existing in the four zones (original soft tissue, uncalcified fibrocartilage, calcified fibrocartilage, and bone) of each kind of insertion, respectively. It then discusses the microstructure constructed from collagen, glycosaminoglycans (GAGs), minerals and others, which provides key support for the biomechanical properties and affects its physiological functions. Finally, the review continues by describing variations in mechanical properties at the millimeter, micrometer, and nanometer scale, which minimize stress concentrations and control stretch at the insertion. In summary, investigating the contrasts between the three has enlightening significance for future directions of repair strategies of insertion diseases and for bioinspired approaches to effective soft-hard interfaces and other tough and robust materials in medicine and engineering.

In vitro synthesis of tensioned synoviocyte bioscaffolds for meniscal fibrocartilage tissue engineering.

BACKGROUND: Meniscal injury is a common cause of lameness in the dog. Tissue engineered bioscaffolds may be a treatment option for meniscal incompetency, and ideally would possess meniscus- like extracellular matrix (ECM) and withstand meniscal tensile hoop strains. Synovium may be a useful cell source for meniscal tissue engineering because of its natural role in meniscal deficiency and its in vitro chondrogenic potential. The objective of this study is to compare meniscal -like extracellular matrix content of hyperconfluent synoviocyte cell sheets ("HCS") and hyperconfluent synoviocyte sheets which have been tensioned over wire hoops (tensioned synoviocyte bioscaffolds, "TSB") and cultured for 1 month. RESULTS: Long term culture with tension resulted in higher GAG concentration, higher chondrogenic index, higher collagen concentration, and type II collagen immunoreactivity in TSB versus HCS. Both HCS and TSB were immunoreactive for type I collagen, however, HCS had mild, patchy intracellular immunoreactivity while TSB had diffuse moderate immunoreactivity over the entire bisocaffold. The tissue architecture was markedly different between TSB and HCS, with TSB containing collagen organized in bands and sheets. Both HCS and TSB expressed alpha smooth muscle actin and displayed active contractile behavior. Double stranded DNA content was not different between TSB and HCS, while cell viability decreased in TSB. CONCLUSIONS: Long term culture of synoviocytes with tension improved meniscal- like extra cellular matrix components, specifically, the total collagen content, including type I and II collagen, and increased GAG content relative to HCS. Future research is warranted to investigate the potential of TSB for meniscal tissue engineering.

Inhibition of CX3CL1 by treadmill training prevents osteoclast-induced fibrocartilage complex resorption during TBI healing.

Introduction: The healing of tendon-bone injuries is very difficult, often resulting in poor biomechanical performance and unsatisfactory functional recovery. The tendon-bone insertion has a complex four distinct layers structure, and previous studies have often focused on promoting the regeneration of the fibrocartilage layer, neglecting the role of its bone end repair in tendon-bone healing. This study focuses on the role of treadmill training in promoting bone regeneration at the tendon-bone insertion and its related mechanisms. Methods: After establishing the tendon-bone insertion injury model, the effect of treadmill training on tendon-bone healing was verified by Micro CT and HE staining; then the effect of CX3CL1 on osteoclast differentiation was verified by TRAP staining and cell culture; and finally the functional recovery of the mice was verified by biomechanical testing and behavioral test. Results: Treadmill training suppresses the secretion of CX3CL1 and inhibits the differentiation of local osteoclasts after tendon-bone injury, ultimately reducing osteolysis and promoting tendon bone healing. Discussion: Our research has found the interaction between treadmill training and the CX3CL1-C3CR1 axis, providing a certain theoretical basis for rehabilitation training.

On how nucleus-endplate integration is achieved at the fibrillar level in the ovine lumbar disc.

The intervertebral disc nucleus has traditionally been viewed as a largely unstructured amorphous gel having little obvious integration with the cartilaginous endplates (CEPs). However, recent work by the present authors has provided clear evidence of structural cohesion across the nucleus-endplate junction via a distinctive microanatomical feature termed insertion nodes. The aim of this study was to explore the nature of these insertion nodes at the fibrillar level. Specially prepared vertebra-nucleus-vertebra composite samples from ovine lumbar motion segments were extended axially and chemically fixed in this stretched state, and then decalcified. Sections taken from the samples were prepared for examination by scanning electron microscopy. A close morphological correlation was obtained between previously published optical microscopic images of the nodes and those seen using low magnification SEM. Progressively high magnifications provided insight into the fibrillar-level modes of structural integration across the nucleus-endplate junction. The closely packed fibrils of the CEP were largely parallel to the vertebral endplate and formed a dense, multi-layer substrate within which the nodal fibrils appeared to be anchored. Our idealised structural model proposes a mechanism by which this integration is achieved. The nodal fibrils, in curving into the CEP, are locked in place within its close-packed layers of transversely aligned fibrils, and probably at multiple levels. Secondly, there appears to be a subtle interweaving of the strongly aligned nodal fibrils with the multi-directional endplate fibrils. It is suggested that this structural integration provides the nucleus with a form of tethered mobility that supports physiological functions quite distinct from the primary strength requirements of the disc.

Tissue-engineering strategies for the tendon/ligament-to-bone insertion.

Injuries to connective tissues are painful and disabling and result in costly medical expenses. These injuries often require reattachment of an unmineralized connective tissue to bone. The uninjured tendon/ligament-to-bone insertion (enthesis) is a functionally graded material that exhibits a gradual transition from soft tissue (i.e., tendon or ligament) to hard tissue (i.e., mineralized bone) through a fibrocartilaginous transition region. This transition is believed to facilitate force transmission between the two dissimilar tissues by ameliorating potentially damaging interfacial stress concentrations. The transition region is impaired or lost upon tendon/ligament injury and is not regenerated following surgical repair or natural healing, exposing the tissue to risk of reinjury. The need to regenerate a robust tendon-to-bone insertion has led a number of tissue engineering repair strategies. This review treats the tendon-to-bone insertion site as a tissue structure whose primary role is mechanical and discusses current and emerging strategies for engineering the tendon/ligament-to-bone insertion in this context. The focus lies on strategies for producing mechanical structures that can guide and subsequently sustain a graded tissue structure and the associated cell populations.

A fresh look at the nucleus-endplate region: new evidence for significant structural integration.

The disc nucleus is commonly thought of as a largely unstructured gel. However, exactly how the nucleus integrates structurally with the endplates remains somewhat ambiguous. The purpose of this study was to investigate whether a substantial level of structural/mechanical cohesion does, in fact, exist across the nucleus-endplate junction. Vertebra-nucleus-vertebra samples were obtained from mature ovine lumbar motion segments and subjected to a novel technique involving circumferential transverse severing (i.e. ring-severing) of the annulus fibrosus designed to eliminate its strain-limiting influence. These samples were loaded in tension and then chemically fixed in order to preserve the stretched nucleus material. Structural continuity across the nucleus-endplate junctions was sufficient for the samples to support, on average, 20 N before tensile failure occurred. Microscopic examination revealed nucleus fibres inserting into the endplates and the significant level of load carried by the nucleus material indicates that there is some form of structural continuity from vertebra to vertebra in the central nucleus region.