Nonwoven-based gelatin/polycaprolactone membrane loaded with ERK inhibitor U0126 for treatment of tendon defects

Background Tendon is a major component of musculoskeletal system connecting the muscles to the bone. Tendon injuries are very common orthopedics problems leading to impeded motion. Up to now, there still lacks effective treatments for tendon diseases. Methods Tendon stem/progenitor cells (TSPCs) were isolated from the patellar tendons of SD rats. The expression levels of genes were evaluated by quantitative RT-PCR. Immunohistochemistry staining was performed to confirm the presence of tendon markers in tendon tissues. Bioinformatics analysis of data acquired by RNA-seq was used to find out the differentially expressed genes. Rat patellar tendon injury model was used to evaluate the effect of U0126 on tendon injury healing. Biomechanical testing was applied to evaluate the mechanical properties of newly formed tendon tissues. Results In this study, we have shown that ERK inhibitor U0126 rather PD98059 could effectively increase the expression of tendon-related genes and promote the tenogenesis of TSPCs in vitro. To explore the underlying mechanisms, RNA sequencing was performed to identify the molecular difference between U0126-treated and control TSPCs. The result showed that GDF6 was significantly increased by U0126, which is an important factor of the TGFβ superfamily regulating tendon development and tenogenesis. In addition, NBM (nonwoven-based gelatin/polycaprolactone membrane) which mimics the native microenvironment of the tendon tissue was used as an acellular scaffold to carry U0126. The results demonstrated that when NBM was used in combination with U0126, tendon healing was significantly promoted with better histological staining outcomes and mechanical properties. Conclusion Taken together, we have found U0126 promoted tenogenesis in TSPCs through activating GDF6, and NBM loaded with U0126 significantly promoted tendon defect healing, which provides a new treatment for tendon injury.

The Ability and Mechanism of nHAC/CGF in Promoting Osteogenesis and Repairing Mandibular Defects

Nano-hydroxyapatite/collagen (nHAC) is a new type of bone tissue engineering scaffold material. To speed up the new bone formation of nHAC, this study used concentrated growth factor (CGF) and nHAC in combination to repair rabbit mandibular defects. nHAC/CGF and nHAC were implanted into rabbit mandibles, and X-ray, Micro-CT, HE and Masson staining, immunohistochemical staining and biomechanical testing were performed at 8, 16 and 24 weeks after surgery. The results showed that as the material degraded, the rate of new bone formation in the nHAC/CGF group was better than that in the nHAC group. The results of the HE and Masson staining showed that the bone continuity or maturity of the nHAC/CGF group was better than that of the nHAC group. Immunohistochemical staining showed that OCN expression gradually increased with time. The nHAC/CGF group showed significantly higher BMP2 than the nHAC group at 8 weeks and the difference gradually decreased with time. The biomechanical test showed that the compressive strength and elastic modulus of the nHAC/CGF group were higher than those of the nHAC group. The results suggest that nHAC/CGF materials can promote new bone formation, providing new ideas for the application of bone tissue engineering scaffold materials in oral clinics.

Standardized fracture creation in the distal humerus and the olecranon for surgical training and biomechanical testing

Introduction Surgical training and biomechanical testing require models that realistically represent the in vivo injury condition. The aim of this work was to develop and test a method for the generation of distal humerus fractures and olecranon fractures in human specimens, while preserving the soft tissue envelope. Methods Twenty-one cadaveric upper extremity specimens (7 female, 14 male) were used. Two different experimental setups were developed, one to generate distal humerus fractures and one to generate olecranon fractures. Specimens were placed in a material testing machine and fractured with a predefined displacement. The force required for fracturing and the corresponding displacement were recorded and the induced energy was derived of the force–displacement graphs. After fracturing, CT imaging was performed and fractures were classified according to the AO classification. Results Eleven distal humerus fractures and 10 olecranon fractures with intact soft tissue envelope could be created. Distal humerus fractures were classified as AO type C (n = 9) and as type B (n = 2), all olecranon fractures were classified as AO type B (n = 10). Distal humerus fractures required significantly more load than olecranon fractures (6077 N ± 1583 vs 4136 N ± 2368, p = 0.038) and absorbed more energy until fracture than olecranon fractures (17.8 J ± 9.1 vs 11.7 J ± 7.6, p = 0.11), while the displacement at fracture was similar (5.8 mm ± 1.6 vs 5.9 mm ± 3.1, p = 0.89). Conclusion The experimental setups are suitable for generating olecranon fractures and distal humerus fractures with intact soft tissue mantle for surgical training and biomechanical testing.

Determination of fighting styles of qualified veteran boxers based on cluster analysis of biomechanical and psychophysiological indicators

Purpose: to reveal the styles of fighting veteran boxers on the basis of a multivariate analysis of psychophysiological and biomechanical indicators. Material and methods. The study involved 42 qualified veteran boxers (age 45-50 years). As research methods, we used a biomechanical analysis of the indicators of the speed of movement of various points and the values ​​of the angles in the joints when performing a direct blow by boxers. The psychophysiological method was used to determine the time of a simple and complex reaction under standard conditions and in various testing modes. We used the method of cluster analysis to distribute athletes into groups using the SPSS - 17.0 program. Within the groups, the athletes are as similar as possible to each other in terms of the analyzed indicators, and between the groups they differ as much as possible. The analysis of the groups of athletes obtained with the help of cluster analysis made it possible to identify athletes with the following styles of fighting: tempo, game, power. Results. Cluster analysis of psychophysiological and biomehanical testing showed the presence of 3 groups of athletes. The clusters were named as follows: Cluster 1 - "Speed and coordination endurance", corresponds to the boxers of the pace of the fight; Cluster 2 - "Speed", corresponds to the boxers of the game style of fighting; Cluster 3 - "Strength and speed", corresponds to the boxers of the pace of the fight. Biomechanical features of boxers of different styles of fighting are reflected in the trajectories of the points of the fist, elbow, knee. Conclusions. The results of this study should be used when planning the individual training of athletes in boxing and to determine the optimal style of competitive competition for qualified veteran boxers. The proposed methods of psychophysiological and biomechanical testing to determine the individual characteristics of boxers are an effective, fairly accessible and convenient tool for revealing the predisposition of boxers to a certain style of fighting.

Hydrogel contained valproic acid accelerates bone-defect repair via activating Notch signaling pathway in ovariectomized rats

The purpose was to observe whether valproic acid (VPA) has a positive effect on bone-defect repair via activating the Notch signaling pathway in an OVX rat model. The MC3T3-E1 cells were cocultured with VPA and induced to osteogenesis, and the osteogenic activity was observed by alkaline phosphatase (ALP) staining, Alizarin Red (RES) staining and Western blotting (WB). Then the hydrogel-containing VPA was implanted into the femoral epiphysis bone-defect model of ovariectomized (OVX) rats for 12 weeks. Micro-CT, biomechanical testing, histology, immunofluorescence, RT-qPCR, and WB analysis were used to observe the therapeutic effect and explore the possible mechanism. ALP and ARS staining and WB results show that the cell mineralization, osteogenic activity, and protein expression of ALP, OPN, RUNX-2, OC, Notch 1, HES1, HEY1, and JAG1 of VPA group is significantly higher than the control group. Micro-CT, biomechanical testing, histology, immunofluorescence, and RT-qPCR evaluation show that group VPA presented the stronger effect on bone strength, bone regeneration, bone mineralization, higher expression of VEGFA, BMP-2, ALP, OPN, RUNX-2, OC, Notch 1, HES1, HEY1, and JAG1 of VPA when compared with OVX group. Our current study demonstrated that local treatment with VPA could stimulate repair of femoral condyle defects, and these effects may be achieved by activating Notch signaling pathway and acceleration of blood vessel and bone formation.

Home-Based Therapy After Stroke Using the Hand Spring Operated Movement Enhancer (HandSOME II)

We have developed a passive and lightweight wearable hand exoskeleton (HandSOME II) that improves range of motion and functional task practice in laboratory testing. For this longitudinal study, we recruited 15 individuals with chronic stroke and asked them to use the device at home for 1.5 h per weekday for 8 weeks. Subjects visited the clinic once per week to report progress and troubleshoot problems. Subjects were then given the HandSOME II for the next 3 months, and asked to continue to use it, but without any scheduled contact with the project team. Clinical evaluations and biomechanical testing was performed before and after the 8 week intervention and at the 3 month followup. EEG measures were taken before and after the 8 weeks of training to examine any recovery associated brain reorganization. Ten subjects completed the study. After 8 weeks of training, functional ability (Action Research Arm Test), flexor tone (Modified Ashworth Test), and real world use of the impaired limb (Motor Activity Log) improved significantly (p < 0.05). Gains in real world use were retained at the 3-month followup (p = 0.005). At both post-training and followup time points, biomechanical testing found significant gains in finger ROM and hand displacement in a reaching task (p < 0.05). Baseline functional connectivity correlated with gains in motor function, while changes in EEG functional connectivity paralleled changes in motor recovery. HandSOME II is a low-cost, home-based intervention that elicits brain plasticity and can improve functional motor outcomes in the chronic stroke population.

New Frontiers towards Regeneration of the Intervertebral Disc: On Progenitor Cells, Growth Factors and Biomaterials

This Special Issue on intervertebral disc (IVD) regeneration focuses on novel advances in understanding the cell sources and culture conditions of various cell types, i.e., progenitor and IVD cells. The issue consists of seven articles that provide a comprehensive overview of recently applied research insights: (1) into how IVD herniation can be provoked in a controlled in vitro biomechanical testing set-up, (2) how cells can be used for IVD repair, (3) the physiological conditions of IVD cells and (4) how hyaluronic acid could be used for IVD repair, and (5) how nucleus pulposus progenitor cells (NPPCs) and mesenchymal stromal cells (MSCs) shall be cultured and expanded towards a possible cell therapy.

The Role of Indian Hedgehog Signaling in Tendon Response to Subacromial Impingement: Evaluation Using a Mouse Model

Background: The underlying cellular and molecular mechanisms involved in the development of tendinopathy due to subacromial supraspinatus tendon (SST) impingement and the response to subsequent removal of impingement remain unknown. Purpose: To investigate the involvement of Indian hedgehog (IHH) signaling in the development of SST tendinopathy and the subsequent healing process after the relief of subacromial impingement in a novel mouse shoulder impingement model. Study Design: Controlled laboratory study. Methods: A total of 48 male wild-type C57BL/6 mice were used in this study. Supraspinatus tendinopathy was induced by inserting a microsurgical clip into the subacromial space bilaterally. Eleven mice were sacrificed at 4 weeks after surgery to establish impingement baseline; 24 mice underwent clip removal at 4 weeks after surgery and then were euthanized at 2 or 4 weeks after clip removal. Thirteen mice without surgical intervention were utilized as the control group. All SSTs were evaluated with biomechanical testing; quantitative histomorphometry after staining with hematoxylin and eosin, Alcian blue, and picrosirius red; and immunohistochemical staining (factor VIII, IHH, Patched1 [PTCH1], and glioma-associated oncogene homolog 1 [GLI1]). Results: The mean failure force and stiffness in the 4-week impingement group decreased significantly compared with the control group ( P < .001) and gradually increased at 2 and 4 weeks after clip removal. Histological analysis demonstrated increased cellularity and disorganized collagen fibers in the SST, with higher modified Bonar scores at 4 weeks, followed by gradual improvement after clip removal. The IHH-positive area and PTCH1- and GLI1-positive cell percentages significantly increased after 4 weeks of clip impingement (20.64% vs 2.06%, P < .001; 53.9% vs 28.03%, P = .016; and 30% vs 12.19%, P = .036, respectively) and continuously increased after clip removal. Conclusion: The authors’ findings suggest that the hedgehog signaling pathway and its downstream signaling mediator and target GLI1 may play a role in the development and healing process of rotator cuff tendinopathy due to extrinsic rotator cuff impingement. Clinical Relevance: This study suggests the potential for the hedgehog pathway, together with its downstream targets, as candidates for further study as potential therapeutic targets in the treatment of supraspinatus tendinopathy.

A Novel and Inexpensive Technique for High-tension Tendon Clamping with Expandable Mesh Sleeving for in Vitro Foot Biomechanics Testing

In vitro biomechanical testing is common in the field of orthopedics when novel devices are investigated prior to human trials. It is typically necessary to apply loads through tendons to simulate normal activities, such as walking during a foot and ankle study. However, attachment of tendons to linear actuators has proven challenging because of the tendency of clamps to either slip off or rupture the tendon. Freeze clamping is generally accepted as the gold standard for very high load testing in excess of 3000 N, but is expensive, time-consuming, and requires significant ancillary equipment. Purely mechanical solutions such as metal jaw clamps, wire meshes, and others have been explored, but these techniques are either costly, have low load capacities, or have not proven to be reproducible. We have developed a novel tendon clamping technique that utilizes a slip-resistant polyester mesh sleeving that encases the tendon and is fixated at the bottom of the tendon/sleeve interaction with a giftbox suture. The loose end of the sleeving can then be tied in to the linear actuator or load cell apparatus using a timber hitch knot. The sleeving technique allows for loads of 2000-2500 N on the Achilles tendon, and is inexpensive, reproducible, and can be modified to apply loads to smaller tendons as well, though a length of tendon/sleeve overlap of at least 16 cm is required to reach maximum loads. This technique should assist researchers in integrating muscle forces into future biomechanical study designs.

Investigation of the effects of two-, four-, six- and eight-strand suture repairs on the biomechanical properties of canine gastrocnemius tenorrhaphy constructs

OBJECTIVE To determine the effects of 2-, 4-, 6- and 8-strand suture repairs on the biomechanical properties of canine gastrocnemius tenorrhaphy constructs in an ex vivo model. SAMPLE 56 cadaveric gastrocnemius musculotendinous units from 28 adult large-breed dogs. PROCEDURES Tendons were randomly assigned to 4 repair groups (2-, 4-, 6- or 8-strand suture technique; n = 14/group). Following tenotomy, repairs were performed with the assigned number of strands of 2-0 polypropylene suture in a simple interrupted pattern. Biomechanical testing was performed. Yield, peak, and failure loads, the incidence of 1- and 3-mm gap formation, forces associated with gap formation, and failure modes were compared among groups. RESULTS Yield, peak, and failure forces differed significantly among groups, with significantly greater force required as the number of suture strands used for tendon repair increased. The force required to create a 1- or 3-mm gap between tendon ends also differed among groups and increased significantly with number of strands used. All constructs failed by mode of suture pull-through. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that increasing the number of suture strands crossing the repair site significantly increases the tensile strength of canine gastrocnemius tendon repair constructs and their resistance to gap formation. Future studies are needed to assess the effects of multistrand suture patterns on tendon glide function, blood supply, healing, and long-term clinical function in dogs to inform clinical decision-making.