Effect of repeated sciatic nerve crush on the conditioning lesion response: Generating an experimental animal model to prolong the denervation period while maintaining peripheral nerve continuity.

Peripheral nerves exhibit long-term residual motor dysfunction following injury. The length of the denervation period before nerve and muscle reconnection is an important factor in motor function recovery. We aimed to investigate whether repeated nerve crush injuries to the same site every 7 days would preserve the conditioning lesion (CL) response and to determine the number of nerve crush injuries required to create an experimental animal model that would prolong the denervation period while maintaining peripheral nerve continuity. Rats were grouped according to the number of sciatic nerve crushes. A significant decrease in the soleus muscle fiber cross-sectional area was observed with increased crushes. After a single crush, macrophage accumulation and macrophage chemotaxis factor CCL2 expression in dorsal root ganglia were markedly increased, which aligned with the gene expression of Ccl2 and its receptor Ccr2. Macrophage numbers, histological CCL2 expression, and Ccl2 and Ccr2 gene expression levels decreased, depending on the number of repeated crushes. Histological analysis and gene expression analysis in the group with four repeated crushes did not differ significantly when compared with uninjured animals. Our findings indicated that repeated nerve crushes at the same site every 7 days sustained innervation loss and caused a loss of the CL response. The experimental model did not require nerve stump suturing and is useful for exploring factors causing prolonged denervation-induced motor dysfunction. SIGNIFICANCE STATEMENT: This study elucidates the effects of repeated nerve crush injury to the same site on innervation and conditioning lesion responses and demonstrates the utility of an experimental animal model that recapitulates the persistent residual motor deficits owing to prolonged denervation without requiring nerve transection and transection suturing.

Chronic Ankle Joint Instability Induces Ankle Sensorimotor Dysfunction: A Controlled Laboratory Study.

BACKGROUND: Chronic ankle instability (CAI) is a clinical sequela that causes the recurrence of ankle sprain by inducing ankle sensorimotor dysfunction. Animal models of CAI have recently shown that ankle ligament injuries mimicking an ankle sprain result in chronic loss of ankle sensorimotor function. However, the underlying mechanisms determining the pathogenesis of CAI remain unclear. HYPOTHESIS: Ankle instability after an ankle sprain leads to the degeneration of the mechanoreceptors, resulting in ankle sensorimotor dysfunction and the development of CAI. STUDY DESIGN: Controlled laboratory study. METHODS: Four-week-old male Wistar rats (N = 30) were divided into 2 groups: (1) the ankle joint instability (AJI) group with ankle instability induced by transecting the calcaneofibular ligament (n = 15) and (2) the sham group (n = 15). Ankle instability was assessed using the anterior drawer test and the talar tilt test at 4, 6, and 8 weeks after the operation (n = 5, for each group at each time point), and ankle sensorimotor function was assessed using behavioral tests, including ladder walking and balance beam tests, every 2 weeks during the postoperative period. Morphology and number of mechanoreceptors in the intact anterior talofibular ligament (ATFL) were histologically analyzed by immunofluorescence staining targeting the neurofilament medium chain and S100 proteins at 4, 6, and 8 weeks postoperatively (n = 5 per group). Sensory neurons that form mechanoreceptors were histologically analyzed using immunofluorescence staining targeting the mechanosensitive ion channel PIEZO2 at 8 weeks postoperatively (n = 5). RESULTS: Ankle sensorimotor function decreased over time in the AJI group, exhibiting decreased ankle instability compared with the sham group (P = .045). The number of mechanoreceptors in the ATFL was reduced (P < .001) and PIEZO2 expression in the sensory neurons decreased (P = .008) at 8 weeks postoperatively. The number of mechanoreceptors was negatively correlated with ankle sensorimotor dysfunction (P < .001). CONCLUSION: The AJI model demonstrated degeneration of the mechanoreceptors in the ATFL and decreased mechanosensitivity of the sensory neurons, which may contribute to CAI. CLINICAL RELEVANCE: Ankle instability causes degeneration of mechanoreceptors and decreases the mechanosensitivity of sensory neurons involved in the development of CAI. This finding emphasizes the importance of controlling ankle instability after ankle sprains to prevent recurrence and the onset of CAI.

The infrapatellar fat pad contributes to spontaneous healing after complete anterior cruciate ligament injury.

Anterior cruciate ligament (ACL) injuries have a very low healing capacity but have recently been shown to heal spontaneously with conservative treatment. This study examined the mechanism of spontaneous ACL healing by focusing on the intra-articular tissues of the knee joint. Skeletally mature Wistar rats (n = 70) were randomly assigned to two groups: the controlled abnormal movement (CAM) and anterior cruciate ligament transection (ACLT) groups. The ACL was completely transected at the mid-portion in both groups. Only the CAM group underwent extra-articular braking to control for abnormal tibial translation. The animals were allowed full cage activity until sacrifice for histological, and molecular biology analyses. The results showed that the behavior of the stump after ACL injury differed between models 12 h after injury. The femoral stump in the ACLT group retreated posteriorly and upwardly. Macrophage polarity analysis revealed that the stump immune response in the CAM group was more activated than that in the ACLT group 6 h after injury. Microarray analysis of the ACL parenchyma and infrapatellar fat pads suggested the involvement of nuclear factor kappa B (NF-κB) signaling. Real-time polymerase chain reaction (PCR) analysis showed that NF-κB gene expression in the infrapatellar fat pad was significantly increased in the CAM group than in the ACLT group. However, there was no difference in the gene expression levels in the ACL parenchyma between models. In conclusion, the healing response of the ACL was activated within 12 h of injury, resulting in differences in the healing response between the models. It has been suggested that infrapatellar fat pads are involved in the healing process and that angiogenesis and antiapoptotic effects through NF-κB signaling may contribute to this mechanism.

The effect of orthosis management on joint instability in knee joint disease: A systematic review.

INTRODUCTION: Joint instability is a common finding of clinical importance in patients with knee disease. This literature review aimed to examine the evidence regarding the effect of orthosis management on joint instability in knee joint disease. METHODS: The detailed protocol for this study was published in the International Prospective Register of Systematic Reviews in the field of health and social welfare (CRD 42022335360). A literature search was conducted on May 2023, using the following databases: Cochrane Central Register of Controlled Trials (CENTRAL), PubMed, Physiotherapy Evidence Database (PEDro), and Institute of Electrical and Electronics Engineers (IEEE) Xplore. A secondary search was manually conducted using Google Scholar to address publication bias. Each database search strategy was described, and the search was conducted by independent reviewers. RESULTS: A total of 281 studies were retrieved, 11 articles were included in the systematic review. Of the 11 articles selected, the number of included diseases was 2 for osteoarthritis, 7 for anterior cruciate ligament injuries, and 3 for posterior cruciate ligament injuries. In result, orthosis management may improve self-reported instability and functional assessment in patients with osteoarthritis, anterior cruciate ligament injury, and posterior cruciate ligament injury. However, an objective evaluation of anatomical instability did not indicate an improvement in joint instability. CONCLUSION: The effects of orthosis management on knee instability might improve physical function and self-reported instability.

Mild treadmill exercise inhibits cartilage degeneration via macrophages in an osteoarthritis mouse model.

Objective: We previously reported how treadmill exercise can suppress cartilage degeneration. Here, we examined the changes in macrophage dynamics in knee osteoarthritis (OA) during treadmill exercise and the effect of macrophage depletion. Design: OA mouse model, generated via anterior cruciate ligament transection (ACLT), was subjected to treadmill exercise of different intensities to investigate the effects on cartilage and synovium. In addition, clodronate liposomes, which deplete macrophages, were injected intra-articularly into the joint to examine the role of macrophages during treadmill exercise. Results: Cartilage degeneration was delayed by mild exercise, and concomitantly, an increase in anti-inflammatory factors in the synovium was observed, with a decrease in the M1 and increase in M2 macrophage ratio. On the contrary, high-intensity exercise led to the progress of cartilage degeneration and was associated with an increase in the M1 and a decrease in the M2 macrophage ratio. The clodronate liposome-induced reduction of synovial macrophages delayed cartilage degeneration. This phenotype was reversed by simultaneous treadmill exercise. Conclusions: Treadmill exercise, especially at high intensity, was detrimental to articular cartilage, whereas mild exercise reduced cartilage degeneration. Moreover, M2 macrophage response appeared necessary for the chondroprotective effect of treadmill exercise. This study indicates the importance of a more comprehensive analysis of the effects of treadmill exercise, not limited to the mechanical stress added directly to cartilage. Hence, our findings might help determine the type and intensity of prescribed exercise therapy for patients with knee OA.

Ankle instability as a prognostic factor associated with the recurrence of ankle sprain: A systematic review.

DESIGN: Systematic review using PRISMA guidelines. PURPOSE: To explore Relationship between ankle instability and ankle sprain recurrence in preventing recurrence of ankle sprains and to provide appropriate treatment. METHODS: MEDLINE (the Cochrane Library) and the Physiotherapy Evidence Database (PEDro) were explored using key words related to ankle instability and ankle sprains in for April 2022. According to the inclusion criteria, studies that 1) targeted patients with ankle sprains, 2) assessed ankle instability, and 3) investigated ankle sprain recurrence rates, were extracted. The author names, publication year, patient characteristics, comparison groups, intervention methods, and outcome data (ankle instability and recurrence) were extracted. A correlation analysis between recurrence rate and ankle instability was conducted. In addition, A meta-analysis was performed on the correlation coefficients within each article. RESULTS: Eight studies were extracted from 149 studies. A correlation analysis was conducted on five studies and meta-analysis was on three studies with the same post-intervention follow-up period and the same assessment methods for ankle instability and recurrence rate. Strong positive correlations were found for the same follow-up periods (r = 0.95: 95%CI [0.62-0.99]; 3-month, r = 0.97: 95%CI [0.75-0.10]; 1 year, p < .05). The correlation became stronger as the follow-up period increased. Furthermore, the meta-analysis showed that ankle instability as well as the main symptoms of sprain, such as pain and swelling, tended to be positively correlated with the recurrent rate of ankle sprains. These results suggest that ankle instability is strongly related to recurrence, and the longer the time since onset, the stronger the relationship. CONCLUSIONS: Ankle instability was a prognostic factor associated with recurrence of ankle sprains in patients with ankle sprains. Therefore, ankle instability is one of important factor in preventing recurrence of ankle sprains.

Structural and pathological changes in the enthesis are influenced by the muscle contraction type during exercise.

Mechanical stress is involved in the onset of sports-related enthesopathy. Although the amount of exercise undertaken is a recognized problem during disease onset, changes in muscle contraction type are also involved in the increase in mechanical stress during exercise. This study aimed to clarify the effects of increased mechanical stress associated with muscle contraction type and amount of exercise on enthesis. Twenty mice underwent treadmill exercise, and the muscle contraction type and overall load during exercise were adjusted by varying the angle and speed conditions. Histological analysis was used to the cross-sectional area of the muscle; area of the enthesis fibrocartilage (FC), and expression of inflammation-, degeneration-, and calcification-related factors in the FC area. In addition, the volume and structure of the bone and FC area were examined using microcomputer imaging. Molecular biological analysis was conducted to compare relative expression levels of inflammation and cytokine-related factors in tendons. The Overuse group, which increased the amount of exercise, showed no significant differences in parameters compared to the sedentary mice (Control group). The mice subjected to slow-speed downhill running (Misuse group) showed pathological changes compared to the Control and Overuse groups, despite the small amount of exercise. Thus, the enthesis FC area may be altered by local mechanical stress that would be increased by eccentric muscle contraction rather than by mechanical stress that increases with the overall amount of exercise. Clinical Significance: The muscle contraction type might be more involved in the onset of sports-related enthesopathy rather than the amount of exercise.

Influence of the site of injury on the spontaneous healing response in a rat model of total rupture of the anterior cruciate ligament.

AIM: The healing ability of the anterior cruciate ligament (ACL) injury is very poor; however, it has recently been shown to undergo self-healing with conservative treatments. In this study, we evaluated the influence of the site of injury on the healing process after complete transverse tear of ACL using a rat model. MATERIALS AND METHODS: A total of 58 skeletally mature Wistar rats were randomly assigned to various ACL injury groups: controlled abnormal movement-mid-portion (CAM-MP), controlled abnormal movement-femoral side (CAM-FS), ACL transection-mid-portion (ACLT-MP), or ACL transection-femoral side (ACLT-FS) injury groups. The ACL was completely transected in the mid-portion in the ACLT-MP and CAM-MP groups, and on the femoral side in the ACLT-FS and CAM-FS groups. Both CAM groups underwent extra-articular braking to control for abnormal tibial translation. The animals were allowed full cage activity until sacrifice postoperatively for histological and biomechanical assessment. RESULTS: Significant differences were found in the ratios of residual ligament lengths between the CAM-MP and CAM-FS groups, demonstrating the validity of each model. Spontaneous healing of the injured ACL was observed in the CAM-MP and CAM-FS groups but not in the ACLT-MP and ACLT-FS groups. The mechanical strength of the healing ACL did not differ between the CAM-MP and CAM-FS groups 8 weeks after injury; however, the former had better mechanical strength than the latter 12 weeks after the injury. CONCLUSION: ACL injuries in the mid-portion and on the femoral side may be treated with conservative therapy for spontaneous healing.

Effect of Various Types of Muscle Contraction with Different Running Conditions on Mouse Humerus Morphology.

How various types of muscle contraction during exercises affect bone formation remains unclear. This study aimed to determine how exercises with different muscle contraction types affect bone morphology. In total, 20 mice were used and divided into four groups: Control, Level, Down Slow, and Down. Different types of muscle contraction were induced by changing the running angle of the treadmill. After the intervention, micro-computed tomography (Micro-CT), tartrate-resistant acid phosphatase/alkaline phosphatase (ALP) staining, and immunohistochemical staining were used to analyze the humerus head, tendon-to-bone attachment, and humerus diaphyseal region. Micro-CT found that the volume ratio of the humeral head, the volume of the tendon-to-bone attachment region, and the area of the humeral diaphyseal region increased in the Down group. However, no difference was detected in bone morphology between the Level and Down Slow groups. In addition, histology showed activation of ALP in the subarticular subchondral region in the Down Slow and Down groups and the fibrocartilage region in the tendon-to-bone attachment. Moreover, Osterix increased predominantly in the Down Slow and Down groups.Overall bone morphological changes in the humerus occur only when overuse is added to EC-dominant activity. Furthermore, different type of muscle contractile activities might promote bone formation in a site-specific manner.

Treadmill Exercise after Controlled Abnormal Joint Movement Inhibits Cartilage Degeneration and Synovitis.

Cartilage degeneration is the main pathological component of knee osteoarthritis (OA), but no effective treatment for its control exists. Although exercise can inhibit OA, the abnormal joint movement with knee OA must be managed to perform exercise. Our aims were to determine how controlling abnormal joint movement and treadmill exercise can suppress cartilage degeneration, to analyze the tissues surrounding articular cartilage, and to clarify the effect of treatment. Twelve-week-old ICR mice (n = 24) underwent anterior cruciate ligament transection (ACL-T) surgery on their right knees and were divided into three groups as follows: ACL-T, animals in the walking group subjected to ACL-T; controlled abnormal joint movement (CAJM), and CAJM with exercise (CAJM + Ex) (n = 8/group). Walking-group animals were subjected to treadmill exercise 6 weeks after surgery, including walking for 18 m/min, 30 min/day, 3 days/week for 8 weeks. Safranin-O staining, hematoxylin-eosin staining, and immunohistochemical staining were performed. The OARSI (Osteoarthritis research Society international) score was lower in the CAJM group than in the ACL-T group and was even lower in the CAJM + Ex group. The CAJM group had a lower meniscal injury score than the ACL-T group, and the CAJM + Ex group demonstrated a less severe synovitis than the ACL-T and CAJM groups. The observed difference in the perichondrium tissue damage score depending on the intervention method suggests different therapeutic effects, that normalizing joint motion can solve local problems in the knee joint, and that the anti-inflammatory effect of treadmill exercise can suppress cartilage degeneration.

Impact of Controlling Abnormal Joint Movement on the Effectiveness of Subsequent Exercise Intervention in Mouse Models of Early Knee Osteoarthritis.

OBJECTIVE: Moderate mechanical stress is necessary for preserving the cartilage. The clinician empirically understands that prescribing only exercise will progress osteoarthritis (OA) for knee OA patients with abnormal joint movement. When prescribing exercise for OA, we hypothesized that degeneration of articular cartilage could be further prevented by combining interventions with the viewpoint of normalizing joint movement. DESIGN: Twelve-week-old ICR mice underwent anterior cruciate ligament transection (ACL-T) surgery in their right knee and divided into 4 groups: ACL-T, controlled abnormal joint movement (CAJM), ACL-T with exercise (ACL-T/Ex), CAJM with exercise (CAJM/Ex). Animals in the walking group were subjected to treadmill exercise 6 weeks after surgery, which included walking for 18 m/min, 30 min/d, 3 d/wk for 4 weeks. Joint instability was measured by anterior drawer test, and safranin-O staining and immunohistochemical staining were performed. RESULTS: OARSI (Osteoarthritis Research Society International) score of ACL-T/Ex group showed highest among 4 groups (P < 0.001). And CAJM/Ex group was lower than ACL-T/Ex group. Positive cell ratio of IL-1ß and MMP-13 in CAJM/Ex group was lower than ACL-T/Ex group (P < 0.05). CONCLUSIONS: We found that the state of the intra-articular environment can greatly influence the effect of exercise on cartilage degeneration, even if exercise is performed under the same conditions. In the CAJM/Ex group where joint movement was normalized, abnormal mechanical stress such as shear force and compression force accompanying ACL cutting was alleviated. These findings may highlight the need to consider an intervention to correct abnormal joint movement before prescribing physical exercise in the treatment of OA.