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.

Morfología y Fisiopatología del Fibrocartílago de la ATM. Revisión sistemática / Morphology and Pathophysiology of the TMJ Fibrocartilage. Systemic review

RESUMEN: El objetivo: del presente estudio fue efectuar una revisión actualizada de la morfología, fisiopatología y aspectos moleculares de la capa fibrocartilaginosa de las ATM en humanos. Método: se realizó búsqueda electrónica y manual con los términos MeSH "TMJ" y "fibrocartilage", sin límite de fecha o idioma. Los desenlaces definidos fueron: morfología del fibrocartílago, fisiopatología del fibrocartílago articular, modeladores moleculares del fibrocartílago y deterioro del fibrocartílago. Resultados: Se identificaron 1.071 títulos de artículos que después de las exclusiones, fueron seleccionados 16 a texto completo para la revisión. El fibrocartílago (FC) que cubre las superficies de las ATM es un tejido crítico en el establecimiento y la tasa de progresión de las alteraciones óseas degenerativas, clínicamente sus discontinuidades se asocian a patologías que provocan dolor, ruidos y limitación funcional. Conclusiones: A pesar de la diversidad metodológica, heterogeneidad de objetivos y diferentes características de los estudios incluidos en la revisión, el fibrocartílago de la ATM, como tejido avascular, determina fisiología de baja capacidad reparativa y mayor frecuencia de patología del FC en mujeres en edad fértil, asociado a receptores hormonales. La fisiopatología muestra reacciones inmunitarias que incrementan la acción de MMPs, interleucinas y FNTα, responsables de la degradación de la matriz extracelular, destrucción celular y morfológica del fibrocartílago, que conducen a sintomatología inflamatoria y degenerativa de pacientes que presentan artralgias y sinovitis en las ATM.

ABSTRACT: The aim: of the present study was to carry out an updated review of the morphology, physiopathology and molecular aspects of the TMJ fibrocartilage in humans. Method: electronic/manual search was performed with the MeSH terms "TMJ" and "fibrocartilage", with no date or language limit. The defined outcomes were: morphology of the fibrocartilage, physiopathology of the articular fibrocartilage, fibrocartilage molecular modelers and fibrocartilage deterioration. Results: 1071 articles were identified and after exclusions, 16 full-texts were selected for review. The fibrocartilage (FC) that covers the surfaces of the TMJ is a critical tissue in the establishment and progression rate of degenerative bone diseases; clinically, its discontinuities are associated with pain, noise and functional limitation. Conclusions: In spite of the methodological diversity, heterogeneity of objectives and different characteristics of the patients included in the review, TMJ fibrocartilage, as avascular tissue, determines a physiology of low reparative capacity, observed with greater frequency of FC damage in women of fertile age, associated with a greater amount of hormonal receptors. The physiopathology shows that the immune reactions increase some MMPs, interleukins and FNTα, which are linked to the degradation of the extracellular matrix, FC cellular and morphological destruction and define the inflammatory and degenerative symptomatology of patients who present TMJ arthralgia´s and synovitis.

Preparation of Decellularized Triphasic Hierarchical Bone-Fibrocartilage-Tendon Composite Extracellular Matrix for Enthesis Regeneration.

Tendon to bone (enthesis) rupture, which may cause disability and persistent pain, shows high rate of re-rupture after surgical repair. Tendon or enthesis scaffolds have been widely studied, but few of these materials can recapitulate the tissue continuity. Thus, this study is conducted to prepare a triphasic decellularized bone-fibrocartilage-tendon (D-BFT) composite scaffold. The D-BFT scaffold is developed using a combination of physical, chemical, and enzymatic treatments using liquid nitrogen, Triton-X 100, sodium-dodecyl sulfate, and DNase I, which effectively removes the cell components while preserving the biological composite and microstructure. Moreover, the mechanical properties of D-BFT are highly preserved and similar to those of the human Achilles tendon. Additionally, in vitro, mesenchymal stem cells (MSCs) adhered, proliferated, and infiltrated into the D-BFT scaffold, and MSC differentiation is confirmed by up-regulation of osteogenic-related and tenogenic-related genes. The repair outcomes are explored by applying the D-BFT scaffold in the model of femur-tibia defects in vivo, which shows good repair results. Thus, the D-BFT scaffold developed in this study is a promising graft for enthesis regeneration.

Bone Surface Micro-Topography at Craniofacial Entheses: Insights on Osteogenic Adaptation at Muscle Insertions.

Macroscopic details of the bone-muscle interface are represented by a mosaic of calcified features inclusive of fossae, tuberosities, crests, and ridges. These features are in part of an adaptive osteogenic response to dissipate forces of localized mechanical loading. In an osteoarchaeological or paleontological context, these features are interpreted as "musculoskeletal stress markers" to infer habitual behaviors. Microscopic surveys of bone surface topography of the enthesis can reveal localized osteogenic topologies. These features illustrate the developmental mechanisms that produce these bony forms and contribute to an evidential basis to read these structures. Microscopic osteogenic topographies at sites of gnathic muscle attachments located in the craniofacial skeleton were explored in reference to extrapolated loading vectors in an ontogenetic series of craniofacial skeletons of the primate (Procolobus verus). Epoxy resin replicas of bone surfaces were made, and micro-topographical detail viewed with Scanning Electron Microscope. Osteoclastic bone remodeling was found at entheses associated with presumptive net tensile loading. Mineralized fibrocartilage was present at entheses, associated with presumptive net compressive loading. Collectively, these outcomes suggest that entheses develop through adaptive osteogenic activity in response to differential vectors of local mechanical loading. However, the presence of mineralized fibrocartilage also suggests that proliferative cartilage has a role in the development of bone eminences providing functional processes. This study concludes that the vector of muscle loading at entheses as well as proliferative fibrocartilage is influencing the form of bony eminences in the primate craniofacial skeleton defining functional and species defining morphologies. Anat Rec, 302:2140-2155, 2019. © 2019 American Association for Anatomy.

Autologous Freeze-Dried, Platelet-Rich Plasma Carrying Icariin Enhances Bone-Tendon Healing in a Rabbit Model.

BACKGROUND: Tendon-bone interface (TBI) injuries are common in sports activities. Owing to the limited regenerative ability of the TBI, its functional healing remains a difficulty in clinical practice. Icariin (ICA) provides strong stimulation for osteogenesis. Platelet-rich plasma (PRP) can be used as a carrier for bioactive molecules, although its ability to provide sustained release for such molecules needs improvement. HYPOTHESIS: Freeze-dried PRP (FD-PRP) as a carrier for ICA can provide sustained release of ICA into the tendon-bone (T-B) healing site, thus accelerating T-B healing. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 84 New Zealand rabbits with partial patellectomy in the hindlimb were randomly allocated into 3 different treatments: ICA incorporated with FD-PRP (ICA/FD-PRP), FD-PRP alone (FD-PRP), or saline control (CTL). The rabbit patella-patellar tendon (PP) interfaces were postoperatively harvested at postoperative week 8 or 16 for gross, radiological, histological, and mechanical evaluations. RESULTS: Our results showed that FD-PRP can act as a carrier for sustained release of ICA into the T-B healing site. Macroscopically, no signs of infection or osteoarthritis were shown in the regenerated PP interfaces, and the area of cartilaginous metaplasia in the FD-PRP and ICA/FD-PRP groups at postoperative week 16 was significantly larger than that of the CTL group (P < .05 for all). Radiologically, micro-computed tomography showed that new bone which formed at the healing site in the ICA/FD-PRP group was significantly increased, remodeled, and mineralized in comparison with the CTL group (P < .05 for all). Histologically, the ICA/FD-PRP group exhibited a significant native PP interface, as shown by the enlargement and remodeling of new bone, well-organized collagen fibers, and robust production of proteoglycans in the regenerated fibrocartilage. The mechanical strength of the regenerated PP interface was significantly improved in the ICA/FD-PRP group. Significantly higher failure load and stiffness were shown in the ICA/FD-PRP group compared with the CTL and FD-PRP groups, respectively (P < .05 for all). CONCLUSION: FD-PRP is a suitable sustained-release carrier for ICA, and ICA/FD-PRP can provide sustained release of ICA into the T-B healing site, thus effectively accelerating T-B healing. CLINICAL RELEVANCE: Findings of this study demonstrate the feasibility of using FD-PRP as a carrier for ICA to improve T-B healing and provide a foundation for future clinical application.

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.

The Importance of the Knee Joint Meniscal Fibrocartilages as Stabilizing Weight Bearing Structures Providing Global Protection to Human Knee-Joint Tissues.

The aim of this study was to review aspects of the pathobiology of the meniscus in health and disease and show how degeneration of the meniscus can contribute to deleterious changes in other knee joint components. The menisci, distinctive semilunar weight bearing fibrocartilages, provide knee joint stability, co-ordinating functional contributions from articular cartilage, ligaments/tendons, synovium, subchondral bone and infra-patellar fat pad during knee joint articulation. The meniscus contains metabolically active cell populations responsive to growth factors, chemokines and inflammatory cytokines such as interleukin-1 and tumour necrosis factor-alpha, resulting in the synthesis of matrix metalloproteases and A Disintegrin and Metalloprotease with ThromboSpondin type 1 repeats (ADAMTS)-4 and 5 which can degrade structural glycoproteins and proteoglycans leading to function-limiting changes in meniscal and other knee joint tissues. Such degradative changes are hall-marks of osteoarthritis (OA). No drugs are currently approved that change the natural course of OA and translate to long-term, clinically relevant benefits. For any pharmaceutical therapeutic intervention in OA to be effective, disease modifying drugs will have to be developed which actively modulate the many different cell types present in the knee to provide a global therapeutic. Many individual and combinatorial approaches are being developed to treat or replace degenerate menisci using 3D printing, bioscaffolds and hydrogel delivery systems for therapeutic drugs, growth factors and replacement progenitor cell populations recognising the central role the menisci play in knee joint health.

Autologous living chondrocytes contained in the meniscal matrix play an important role in in vivo meniscus regeneration induced by in situ meniscus fragment implantation.

INTRODUCTION: Implantation of autogenous meniscal fragments wrapped with a fascia sheath significantly enhances fibrocartilage regeneration in vivo in defect cases at 12 weeks after implantation. The specific effects of the implanted autologous living chondrocytes and meniscal matrix have not been elucidated, however. The aim of this study was to clarify the role of autologous living chondrocytes contained in the meniscal matrix in in vivo meniscus regeneration induced by in situ meniscus fragment implantation. HYPOTHESIS: Implantation of meniscus fragments containing autologous living chondrocytes may result in significant in vivo meniscus regeneration. MATERIALS AND METHODS: Seventy-five rabbits were used in this study. A partial meniscectomy of the anterior one-third of the medial meniscus including the part of the anterior horn was performed. The rabbits were divided into 3 groups. In Group I, no treatment was applied to the defect. In Group II, the autogenous meniscal fragments devitalized by freeze-thaw treatment were reimplanted into the defect. In Group III, the autogenous meniscal fragments were reimplanted. In each group, the defect was covered with a fascia. Five rabbits from each group were subjected to morphologic and histologic evaluations at 3, 6, and 12 weeks, and 5 rabbits from each group were subjected to biomechanical evaluations at 6 and 12 weeks. RESULTS: Histologically, no cells were seen in the grafted meniscal fragments at 3 weeks in Group II, whereas chondrocytes in the grafted meniscal fragments were alive at 3 weeks in Group III. Histologic and biomechanical data for Group II were slightly but significantly better than those of Group I at 12 weeks after implantation (p=0.007 and p=0.002, respectively), whereas the data for Group III were significantly superior to those of Groups I and II at 12 weeks (p<0.0014 and p<0.0029, respectively). DISCUSSIONS: Grafted autologous living chondrocytes contained in the meniscal matrix play an important role in in vivo meniscus regeneration induced by in situ meniscus fragment implantation. STUDY DESIGN: II, Controlled laboratory study.

Effect of Low-Intensity Pulsed Ultrasound After Autologous Adipose-Derived Stromal Cell Transplantation for Bone-Tendon Healing in a Rabbit Model.

BACKGROUND: Low-intensity pulsed ultrasound (LIPUS), as a safe biophysiotherapy, can enhance bone-tendon (B-T) healing in vivo and induce osteogenic or chondrogenic differentiation of mesenchymal stromal cells in vitro. This study aimed to determine whether LIPUS can improve the efficacy of transplanted mesenchymal stromal cells on B-T healing. HYPOTHESIS: LIPUS can induce lineage-specific differentiation of transplanted adipose-derived stromal cells (ASCs) at the B-T healing site, thus resulting in superior healing quality when compared with LIPUS or ASCs alone. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 112 mature rabbits with partial patellectomy in the hindlimb were randomly assigned into mock sonication without ASCs (control), ultrasonication without ASCs (LIPUS), mock sonication with ASCs (ASCs), and ultrasonication with ASCs (LIPUS + ASCs). The treatment time of the mock sonication or ultrasonication was 20 minutes per day. Autologous ASCs were transplanted to the healing site by fibrin glue during the operation, and LIPUS was delivered daily starting at postoperative day 3 until euthanasia. The patella-patellar tendon junctions were postoperatively harvested at 8 and 16 weeks for radiological, histological, and mechanical evaluations. Additionally, 9 animals were used for ASC tracking with mCherry protein. RESULTS: Radiologically, there was more new bone formation and remodeling in the LIPUS + ASCs group as compared with the other groups. Synchrotron radiation micro-computed tomography showed that the LIPUS + ASCs group significantly increased bone volume fraction, trabecular thickness, and trabecular number at the healing site as compared with the other groups at postoperative 8 weeks ( P < .05 for all). Histologically, immunohistochemical staining confirmed that the transplanted mCherry-ASCs can differentiate into osteoblasts and fibrochondrocytic-like cells. Meanwhile, as compared with the other groups, the LIPUS + ASCs group showed more formation and maturity of the fibrocartilage layer and new bone at postoperative weeks 8 and 16 ( P < .05 for all). Biomechanically, the LIPUS + ASCs group showed significantly higher failure load and stiffness versus the other groups at postoperative weeks 8 and 16 ( P < .05 for all). CONCLUSION: Autologous ASC transplantation stimulated with LIPUS can result in superior B-T healing quality when compared with LIPUS or ASCs alone. CLINICAL RELEVANCE: This study demonstrates the effectiveness of using ASC transplantation stimulated with LIPUS for B-T healing and provides a foundation for future clinical studies.

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.

Effects of Ultra-Purified Alginate Gel Implantation on Meniscal Defects in Rabbits.

BACKGROUND: Many tissue-engineered methods for meniscal repair have been studied, but their utility remains unclear. HYPOTHESIS: Implantation of low-endotoxin, ultra-purified alginate (UPAL) gel without cells could induce fibrocartilage regeneration on meniscal defects in rabbits. STUDY DESIGN: Controlled laboratory study. METHODS: Forty-two mature Japanese White rabbits were divided into 2 groups of 21 animals each. In each animal, a cylindrical defect measuring 2 mm in diameter was created with a biopsy punch on the anterior horn of the medial meniscus. In the control group, no treatment was applied on the left medial meniscal defect. In the UPAL gel group, the right medial meniscal defect was injected with the UPAL gel and gelated by a CaCl2 solution. Samples were evaluated at 3, 6, and 12 weeks postoperatively. For biomechanical evaluation, 6 additional samples from intact animals were used for comparison. RESULTS: The macroscopic score was significantly greater in the UPAL gel group than in the control group at 3 weeks (mean ± SE: 5.6 ± 0.82 vs 3.4 ± 0.83, P = .010), 6 weeks (5.9 ± 0.72 vs 2.5 ± 0.75, P = .026), and 12 weeks (5.2 ± 1.21 vs 1.0 ± 0.63, P = .020). The histological score was significantly greater in the UPAL group than in the control group at 3 weeks (2.1 ± 0.31 vs 1.2 ± 0.25, P = .029) and 12 weeks (2.2 ± 0.55 vs 0.3 ± 0.21, P = .016). The mean stiffness of the reparative tissue in the UPAL gel group was significantly greater than that in the control group at 6 weeks (24.325 ± 3.920 N/mm vs 8.723 ± 1.190 N/mm, P = .006) and at 12 weeks (27.804 ± 6.169 N/mm vs not applicable [because of rupture]). CONCLUSION: The UPAL gel enhanced the spontaneous repair of fibrocartilage tissues in a cylindrical meniscal defect in rabbits. CLINICAL RELEVANCE: These results imply that the acellular UPAL gel may improve the repair of traumatic meniscal injuries.

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.

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.

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.

Effect of Suture Absorbability on Rotator Cuff Healing in a Rabbit Rotator Cuff Repair Model.

BACKGROUND: Various suture materials can be clinically used for rotator cuff repair (RCR). RCR with high-strength nonabsorbable sutures may not be ideal, because it may cause stress shielding, which may hinder enthesis regeneration and maturation in the tendon-bone interface. RCR with strength-decreasing sutures (ie, absorbable sutures) may be a better choice. However, the effects of suture absorbability on enthesis regeneration and maturation have not been investigated. HYPOTHESIS: The use of absorbable sutures in RCR would produce a better tendon-bone connection structure, which provides histological and biomechanical advantages over the use of nonabsorbable sutures. STUDY DESIGN: Controlled laboratory study. METHODS: A supraspinatus tear was created on the right shoulder in 108 of 120 skeletally mature male rabbits. The animals were randomly divided into 3 groups, with 36 rabbits in each group, to undergo RCR individually with total absorbable, partial absorbable, and nonabsorbable sutures (TAS, PAS, and NAS). Twelve animals in each group were sacrificed at 4, 8, and 12 weeks after surgery, with 6 operated shoulders used for histological evaluation to detect enthesis regeneration and maturation and the other 6 for biomechanical testing. The remaining 12 animals without supraspinatus tear were used as control. RESULTS: At 12 weeks, in the tendon-bone interface, enthesis regeneration was detected in the TAS group but not in the NAS group. A mature enthesis appeared in the TAS group but not in the NAS group. In the PAS group, enthesis regeneration was also observed; however, the fibrocartilage was not abundant and the enthesis maturity not good as compared with the TAS group. Biomechanical testing showed that the rotator cuff-greater tuberosity connection structure in the TAS and PAS groups had greater values of ultimate load to failure, stiffness, and stress than the NAS group at all time points. CONCLUSION: In RCR in an acute rabbit rotator cuff tear model, the use of sutures with absorbability lead to enthesis regeneration, increased maturity of rotator cuff insertion, and enhanced rotator cuff-greater tuberosity connection. CLINICAL RELEVANCE: Compared with the use of NAS, the use of TAS or PAS might be a better choice for RCR.

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.

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.

Hydraulic permeability of meniscus fibrocartilage measured via direct permeation: Effects of tissue anisotropy, water volume content, and compressive strain.

Hydraulic permeability is an important material property of cartilaginous tissues, governing the rate of fluid flow, which is crucial to tissue biomechanics and cellular nutrition. The effects of strain, anisotropy, and region on the hydraulic permeability in meniscus tissue have not been fully elucidated. Using a one-dimensional direct permeation test, we measured the hydraulic permeability within statically compressed porcine meniscus specimens, prepared such that the explants were in either the axial or circumferential direction of either the central or horn (axial direction only) region of the medial and lateral menisci. A constant flow was applied and the pressure difference was measured using pressure transducers. Specimens were tested under 10-20% compressive strain. Permeability values were in the range of 1.53-1.87 × 10-15 m4/Ns, which is comparable to values found in the literature. Permeability was significantly anisotropic, being higher in the circumferential direction than in the axial direction. Additionally, there was a significant negative correlation between strain level and permeability for all groups. Lastly, no statistically significant difference was found between permeability coefficients from different regional locations. This study provides important information regarding structure-function relationships in meniscal tissues that helps to elucidate biomechanics and transport in the tissue, and can aid in the understanding of the tissue's role in the function of the knee joint and onset of osteoarthritis.

A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle.

The temporomandibular joint (TMJ) has the capacity to adapt to external stimuli, and loading changes can affect the position of condyles, as well as the structural and cellular components of the mandibular condylar cartilage (MCC). This manuscript describes methods for analyzing these changes and a method for altering the loading of the TMJ in mice (i.e., compressive static TMJ loading). The structural evaluation illustrated here is a simple morphometric approach that uses the Digimizer software and is performed in radiographs of small bones. In addition, the analysis of cellular changes leading to alterations in collagen expression, bone remodeling, cell division, and proteoglycan distribution in the MCC is described. The quantification of these changes in histological sections - by counting the positive fluorescent pixels using image software and measuring the distance mapping and stained area with Digimizer - is also demonstrated. The methods shown here are not limited to the murine TMJ, but could be used on additional bones of small experimental animals and in other regions of endochondral ossification.

Time-Dependent Changes in the Structure of Calcified Fibrocartilage in the Rat Achilles Tendon-Bone Interface With Sciatic Denervation.

The enthesis transmits a physiological load from soft to hard tissue via fibrocartilage. The histological alterations induced by this physiological loading remain unclear. This study was performed to examine the histomorphological alterations in the collagen fiber bundle alignment and depth of collagen interdigitation between the calcified fibrocartilage and the bone. We examined the Achilles enthesis of rats with sciatic denervation to explore the mechanical effects of structural changes in the enthesis. The parallelism of the collagen fiber bundles was significantly reduced 8 weeks after denervation. However, the depth of collagen interdigitation significantly increased at 2 and 4 weeks after denervation and then significantly decreased 8 weeks after denervation. In conclusion, a lack of muscle loading induced structural alterations in the distal calcified fibrocartilage. These findings suggest that while structural changes in the enthesis are necessary for the development of physiological loading, structural deformities are required in the long term. Anat Rec, 300:2166-2174, 2017. © 2017 Wiley Periodicals, Inc.