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Simple Summary: Programmed death-ligand 1 (PD-L1) on the tumor cell surface binds to its receptor, programmed death-1 (PD-1), on T cells and inhibits their activity. Immunotherapy involving PD-L1 and PD-1 inhibition has provided a remarkable innovation in cancer therapy. Recently, the soluble form of PD-L1 (sPD-L1) has attracted attention for systemic immune suppression. In this study, we successfully showed that sarcoma cells can release functional exosomal PD-L1, one of the forms of sPD-L1. Furthermore, the release of exosomal PD-L1 derived from normal and sarcoma cells was shown to be induced by irradiation. These findings demonstrate that radiated cells, including normal cells and sarcoma cells, induce the systemic release of exosomal PD-L1, and combination therapy with anti-PD-1/PD-L1 antibody may block the immune activity of exosomal PD-L1. (1) Background: Exosomal PD-L1 has garnered attention owing to its role in instigating systemic immune suppression. The objective of this study is to elucidate whether bone and soft tissue sarcoma cells possess the capacity to secrete functionally active exosomal PD-L1 and whether radiotherapy (RT) induces the exosomal PD-L1 release. (2) Methods: Human osteosarcoma cell line 143B and human fibrosarcoma cell line HT1080 were utilized. Exosomes were isolated from the culture medium and blood via ultracentrifugation. The expression of PD-L1 on both tumor cells and exosomes was evaluated. The inhibitory effect on PBMC was employed to assess the activity of exosomal PD-L1. Post radiotherapy, changes in PD-L1 expression were compared. (3) Results: Exosomal PD-L1 was detected in the culture medium of tumor cells but was absent in the culture medium of PD-L1 knockout cells. Exosomal PD-L1 exhibited an inhibitory effect on PBMC activation. In tumor-bearing mice, human-derived exosomal PD-L1 was detected in the bloodstream. Following radiotherapy, tumor cells upregulated PD-L1, and human-derived exosomal PD-L1 were detected in the bloodstream. (4) Conclusions: Exosomal PD-L1 emanates from bone and soft tissue sarcoma cells and is disseminated into the circulatory system. The levels of PD-L1 in tumor cells and the release of exosomal PD-L1 were augmented after irradiation with RT. [ABSTRACT FROM AUTHOR]

Background: This study aimed to report the long-term outcomes of total hip arthroplasty (THA) using a Conserve Plus (Wright Medical, Japan) metal-on-metal (MoM) acetabular prosthesis with a modular neck stem. Methods: This study enrolled 50 patients (10 men and 40 women; mean age, 65.8 (39–87) years) who underwent primary THA using a Conserve Plus MoM acetabular prosthesis with a modular neck stem. The preoperative diagnosis in most patients was osteoarthritis. Clinical function of hip joint outcomes was investigated using the Japanese Orthopedic Association (JOA) hip score preoperatively and at the final follow-up. The perfect JOA hip score was 100, while the worst score was 0. Radiological analyses were evaluated during the final follow-up visit. Magnetic resonance imaging (MRI) images were evaluated to screen for pseudotumors in 43 hips postoperatively. Results: Six patients did not visit before their 10-year follow-up for unknown reasons. Therefore, 44 patients were evaluated at a mean of 11-years of follow-up (10–12 years). The mean (±SD) preoperative JOA hip score of 44.2 (±15.5) improved significantly to 85.1 (±12.9) postoperatively at the final follow-up (n = 36 hips, excluding eight revision cases). One patient underwent femoral fixation for a periprosthetic fracture due to trauma that occurred 4 years postoperatively. Spot welds were identified in 93.2% (41/44 hips) of cases. Severe (third- and fourth-degree) stress shielding was identified in 40.9% (18/44 hips) of cases. Twenty-two patients (51.2%) had pseudotumors attributable to MoM articulation based on MRI results, 2 to 10 years after arthroplasty. Three hips showed cup osteolysis (7%) and three showed trochanteric region osteolysis (7%). There were seven cup and/or three stem revisions for aseptic loosening and/or osteolysis at 4 months (with trauma) and 3.3 to 11 years (with pseudotumor) postoperatively. The Kaplan–Meier survivorship for the THA construct in this group was constant at 93.0% and 75.9% at 10 and 12 years after arthroplasty, respectively. The rates of survivorship of revision and loss of follow-up at 10 and 12 years were 83.9% and 66.8%, respectively. Conclusions: In summary, we reported on the long-term treatment results of MoM THA, precautions based on our cohort's findings, and the measures taken to address these issues, such as revision replacement and its outcomes. Clinical scores revealed good outcomes during the mean 11-year follow-up period. However, the prevalence of pseudotumors (PTs) was 51.2%. Some cases required revisions even after the 10 years following surgery. This is because in MoM THA, PT occurrence increases over time, and as a result, there were cases in which revised THA was required even after 10 years. [ABSTRACT FROM AUTHOR]

Simple Summary: A total of 217 Japanese patients who underwent surgical resection for extremity STS were included. The Sarculator-predicted 10-year probability of overall survival (pr-OS) was stratified into two subgroups: lower risk (10-year pr-OS ≥ 60%) and higher risk (10-year pr-OS < 60%). The modified Glasgow prognostic score (mGPS) varied from 0 to 2. We showed that Sarculator is a validated nomogram designed to predict overall survival. Among the patients with a higher risk, those with an mGPS of 1 or 2 had poorer OS compared to those with a score of 0. The mGPS could potentially play an important role in identifying patients who are at high risk of death and metastasis in the higher-risk group on the Sarculator. Background: Sarculator is a validated nomogram designed to predict overall survival (OS) in extremity soft tissue sarcoma (STS). Inflammation plays a critical role in cancer development and progression. There were no reports which investigated the relationship between Sarculator and inflammation. Methods: A total of 217 patients with extremity STS were included. The Sarculator-predicted 10-year probability of OS (pr-OS) was stratified into two subgroups: lower risk (10-year pr-OS ≥ 60%) and higher risk (10-year pr-OS < 60%). The modified Glasgow prognostic score (mGPS) varied from 0 to 2. Results: Out of the 217 patients, 67 were classified as higher risk, while 150 were lower risk. A total of 181 patients had an mGPS of 0, and 36 had a score of 1 or 2. The 5-year OS was 83.3%. When patients were divided into two groups according to the 10-year pr-OS, those with a higher risk had poorer OS than those with a lower risk. Among the patients with a higher risk, those with an mGPS of 1 or 2 had poorer OS compared to those with a score of 0. Conclusions: The mGPS could potentially play an important role in identifying patients who are at high risk of death and metastasis in the higher-risk group on the Sarculator. [ABSTRACT FROM AUTHOR]

Glial-cell-line-derived neurotrophic factor (GDNF) family ligands (GFLs) contribute to the sensitization of primary afferents and are involved in the pathogenesis of inflammatory pain. The purpose of this preliminary study was to examine the expression of other GFLs (neurturin (NRTN), artemin (ARTN), persephin (PSPN)) and receptors in human IVD cells and tissues exhibiting early and advanced stages of degeneration. Human IVD cells were cultured as a monolayer after isolation from the nucleus pulposus (NP) and anulus fibrosus (AF) tissues. The mRNA expression of NRTN, ARTN, PSPN, and their receptors (GFRA2–GFRA4) was quantified using real-time PCR. Protein expression was evaluated using immunohistochemistry and Western blotting. The expression of NRTN, ARTN, PSPN, and their co-receptors (GFRA2-GFRA4) was identified in human IVD cells at both mRNA and protein levels. A trend was noted wherein the mRNA expression of ARTN, PSPN, and GFRA2 was upregulated by IL-1β treatment in a dose-dependent manner. The percentages of immunopositive cells in the advanced degenerate stage of ARTN, PSPN, and GFRA2 were significantly higher than those in the early degenerate stage. Their expression was enhanced in advanced tissue degeneration, which suggests that GFLs (ARTN and PSPN) may be involved in the pathogenesis of discogenic pain. [ABSTRACT FROM AUTHOR]

This study aimed to quantitatively assess disc bulging using computed tomography (CT) in patients with lumbar spinal stenosis (LSS) and to examine whether disc bulging affects the surgical outcomes of patients with LSS after posterior decompression surgery. Sixty-three patients who underwent posterior decompression surgery for LSS were included. The extent of disc bulging was evaluated as the percentage of the extended area of the disc against the endplate area (%EAD) on axial CT images. The participants completed the following clinical outcome assessments (COAs) preoperatively and 12 months postoperatively: the JOA Back Pain Evaluation Questionnaire (JOABPEQ), Oswestry Disability Index (ODI), and Roland–Morris Disability Questionnaire (RDQ). The mean %EAD of 315 intervertebral discs was 18.9 ± 8.0. The %EAD was highest at L4/L5, followed by L3/L4, L2/L3, L1/L2, and L5/S1. The %EAD of the surgical level showed no significant correlation with all the preoperative COAs, but it had significant correlation with lumbar function, walking ability, social function domains of the JOABPEQ, ODI score, and RDQ score 12 months postoperatively. %EAD was significantly associated with the postoperative score in the walking ability domain of the JOABPEQ. %EAD affects postoperative clinical outcomes, including low back pain-related quality of life after decompression surgery. [ABSTRACT FROM AUTHOR]

Despite its widespread existence, there are relatively few drugs that can inhibit the progression of osteoarthritis (OA). Syndecan-4 (SDC4) is a transmembrane heparan sulfate proteoglycan that modulates cellular interactions with the extracellular matrix. Upregulated SDC4 expression in articular cartilage chondrocytes correlates with OA progression. In the present study, we treated osteoarthritic cartilage with SDC4 to elucidate its role in the disease's pathology. In this in vitro study, we used real-time polymerase chain reaction (PCR) to investigate the effects of SDC4 on anabolic and catabolic factors in cultured chondrocytes. In the in vivo study, we investigated the effect of intra-articular injection of SDC4 into the knee joints of an OA mouse model. In vitro, SDC4 upregulated the expression of tissue inhibitor of metalloproteinase (TIMP)-3 and downregulated the expression of matrix metalloproteinase (MMP)-13 and disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-5 in chondrocytes. Injection of SDC4 into the knee joints of OA model mice prevented articular cartilage degeneration 6 and 8 weeks postoperatively. Immunohistochemical analysis 8 weeks after SDC4 injection into the knee joint revealed decreased ADAMTS-5 expression and increased TIMP-3 expression. The results of this study suggest that the treatment of osteoarthritic articular cartilage with SDC4 inhibits cartilage degeneration. [ABSTRACT FROM AUTHOR]

Background and Objectives: Adult (de novo) degenerative scoliosis (ADS) develops through degenerative changes in the lumbar spine, leading to spinal malalignment, which usually progresses with age. Strong evidence for non-operative care in patients with ADS is lacking, and whether physical exercise can improve the scoliosis curve remains unknown. Materials and Methods: We present a case of early stage ADS in which the coronal imbalance was improved by daily training. A 65-year-old female patient complained of lower back pain (LBP) and bilateral leg pain. She was diagnosed with early stage ADS with lumbar degenerative spondylolisthesis by imaging. She completed six months of daily physical training, including swimming, aerobic bikes, stretching, yoga, and Taijiquan. Results: Her LBP and neurological symptoms improved, and coronal–spinal balance was restored, which was maintained for four years by continued daily physical training. Conclusions: This is the first case of a 65-year-old ADS patient whose coronal balance was significantly restored through daily physical training. Substantial physical training focused on trunk muscle strength is important for spinal stabilization and for improving spinal malalignment in patients with early stage ADS. [ABSTRACT FROM AUTHOR]

Simple Summary: High-grade bone and soft tissue sarcomas are known for high recurrence and low survival rates. New drug development is costly and time-consuming. Instead, new combinations of existing drugs may offer novel anticancer strategies. We previously demonstrated that the combination of the mammalian target of rapamycin inhibitor everolimus and the proteasome inhibitor bortezomib is effective against osteosarcoma cells in a drug screening test. Building on previous research, we showed that the combination of everolimus and bortezomib exerted synergistic antiproliferative and apoptotic effects against fibrosarcoma and osteosarcoma, suppressing pulmonary metastases in osteosarcoma-bearing mice. Furthermore, we identified that everolimus and bortezomib inhibited tumor growth via the JNK/p38/ERK MAPK and AKT pathways. This combination treatment may be effective against bone and soft tissue sarcoma suppressing tumor growth and metastasis. The combination of the mammalian target of rapamycin and proteasome inhibitors is a new treatment strategy for various tumors. Herein, we investigated the synergistic effect of everolimus and bortezomib on tumor growth and metastasis in bone and soft tissue sarcomas. The antitumor effects of everolimus and bortezomib were assessed in a human fibrosarcoma (FS) cell line (HT1080) and mouse osteosarcoma (OS) cell line (LM8) by MTS assays and Western blotting. The effects of everolimus and bortezomib on HT1080 and LM8 tumor growth in xenograft mouse models were evaluated using tumor volume and the number of metastatic nodes of the resected lungs. Immunohistochemistry was used to evaluate cleaved PARP expression. The combination therapy decreased FS and OS cell proliferation compared with either drug alone. This combination induced more intense p-p38, p-JNK, and p-ERK and activated apoptosis signals, such as caspase-3, compared with single-agent treatment. The combination treatment reduced p-AKT and MYC expression, decreased FS and OS tumor volumes, and suppressed lung metastases of OS. The combination therapy inhibited tumor growth in FS and OS and metastatic progression of OS via the JNK/p38/ERK MAPK and AKT pathways. These results could aid in the development of new therapeutic strategies for sarcomas. [ABSTRACT FROM AUTHOR]

We assessed the clinical results of irrigation and debridement (I&D) with antibiotic-impregnated calcium hydroxyapatite (CHA) as a novel antibiotic delivery system for the treatment of prosthetic-joint-associated infection (PJI) after total hip arthroplasty (THA). Thirteen patients (14 hips) treated with I&D for PJI after THA at our institution between 1997 and 2017 were retrospectively evaluated. The study group included four men (five hips) and nine women, with an average age of 66.3 years. Four patients (five hips) had symptoms of infection within less than 3 weeks; however, nine patients had symptoms of infection over 3 weeks. All patients received I&D with antibiotic-impregnated CHA in the surrounding bone. In two hips (two cups and one stem), cup and/or stem revision were performed with re-implantation because of implant loosening. In ten patients (11 hips), vancomycin hydrochloride was impregnated in the CHA. The average duration of follow-up was 8.1 years. Four patients included in this study died of other causes, with an average follow-up of 6.7 years. Eleven of thirteen patients (12 of 14 hips) were successfully treated, and no signs of infection were observed at the latest follow-up. In two patients (two hips) for whom treatment failed, infection was successfully treated with two-stage re-implantation. Both patients had diabetes mellitus and symptoms of infection over 3 weeks. Eighty-six percent of patients were successfully treated. No complications were observed with this antibiotic-impregnated CHA. I&D treatment with antibiotic-impregnated CHA produced a higher rate of success in patients with PJI after THA. [ABSTRACT FROM AUTHOR]

Spinal diseases are commonly associated with pain and neurological symptoms, which negatively impact patients' quality of life. Platelet-rich plasma (PRP) is an autologous source of multiple growth factors and cytokines, with the potential to promote tissue regeneration. Recently, PRP has been widely used for the treatment of musculoskeletal diseases, including spinal diseases, in clinics. Given the increasing popularity of PRP therapy, this article examines the current literature for basic research and emerging clinical applications of this therapy for treating spinal diseases. First, we review in vitro and in vivo studies, evaluating the potential of PRP in repairing intervertebral disc degeneration, promoting bone union in spinal fusion surgeries, and aiding in neurological recovery from spinal cord injury. Second, we address the clinical applications of PRP in treating degenerative spinal disease, including its analgesic effect on low back pain and radicular pain, as well as accelerating bone union during spinal fusion surgery. Basic research demonstrates the promising regenerative potential of PRP, and clinical studies have reported on the safety and efficacy of PRP therapy for treating several spinal diseases. Nevertheless, further high-quality randomized controlled trials would be required to establish clinical evidence of PRP therapy. [ABSTRACT FROM AUTHOR]

Background and Objectives: Recently, the clinical application of platelet-rich plasma (PRP) has gained popularity for the treatment of degenerative disc diseases. However, the regenerative effects and factors associated with treatment outcomes after intradiscal injection of PRP remain unknown. This study aimed to evaluate time-dependent changes in imaging findings related to intervertebral disc (IVD) degeneration and to identify factors associated with the outcomes of PRP injection therapy. Materials and Methods: A retrospective analysis of a previous randomized clinical trial of intradiscal injection of the releasate isolated from PRP (PRPr) in patients with discogenic low back pain (LBP) was performed. Radiographic parameters (segmental angulation and lumbar lordosis) and MRI phenotypes, including Modic changes, disc bulge, and high-intensity zones (HIZs), were evaluated at baseline and 6 and 12 months post-injection. Treatment outcomes were evaluated based on the degree of LBP and LBP-related disability at 12 months post-injection. Results: A total of 15 patients (mean age: 33.9 ± 9.5 years) were included in this study. Radiographic parameters showed no significant changes after the PRPr injection. There were no remarkable changes in the prevalence or type of MRI phenotype. Treatment outcomes were significantly improved after treatment; however, the number of targeted discs and the presence of posterior HIZs at baseline were significantly but negatively associated with treatment outcomes. Conclusions: Intradiscal injection of PRPr significantly improved LBP and LBP-related disability 12 months post-injection; however, patients with multiple target lesions or posterior HIZs at baseline were significantly associated with poor treatment outcomes. [ABSTRACT FROM AUTHOR]

Marked cellular changes occur in human intervertebral disc (IVD) degeneration during disc degeneration with biochemical changes. Genome-wide analysis of the DNA methylation profile has identified 220 differentially methylated loci associated with human IVD degeneration. Among these, two cell-cycle–associated genes, growth arrest and DNA damage 45 gamma (GADD45G) and cytoplasmic activation/proliferation-associated protein-1 (CAPRIN1), were focused on. The expression of GADD45G and CAPRIN1 in human IVDs remains unknown. We aimed to examine the expression of GADD45G and CAPRIN1 in human nucleus pulposus (NP) cells and evaluate those in human NP tissues in the early and advanced stages of degeneration according to Pfirrmann magnetic resonance imaging (MRI) and histological classifications. Human NP cells were cultured as monolayers after isolation from NP tissues by sequential enzyme digestion. Total RNA was isolated, and the mRNA expression of GADD45G and CAPRIN1 was quantified using real-time polymerase chain reaction. To examine the effects of pro-inflammatory cytokines on mRNA expression, human NP cells were cultured in the presence of IL-1β. Protein expression was evaluated using Western blotting and immunohistochemistry. GADD45G and CAPRIN1 expression was identified in human NP cells at both mRNA and protein levels. The percentage of cells immunopositive for GADD45G and CAPRIN1 significantly increased according to the Pfirrmann grade. A significant correlation between the histological degeneration score and the percentage of GADD45G-immunopositive cells was identified, but not with that of CAPRIN1-immunopositive cells. The expression of cell-cycle-associated proteins (GADD45G and CAPRIN1) was enhanced in human NP cells at an advanced stage of degeneration, suggesting that it may be regulated during the progression of IVD degeneration to maintain the integrity of human NP tissues by controlling cell proliferation and apoptosis under epigenetic alteration. [ABSTRACT FROM AUTHOR]

Ultra-high-molecular-weight polyethylene (UHMWPE) wear and particle-induced osteolysis contribute to the failure of total hip arthroplasty (THA) and total knee arthroplasty (TKA). Highly crosslinked polyethylene (HXLPE) was developed in the late 1990s to reduce wear and has shown lower wear rates and loosening than conventional UHMWPE in THA. The irradiation dose for crosslinking is up to 100 kGy. However, during crosslinking, free radical formation induces oxidation. Using HXLPE in THA, the cumulative revision rate was determined to be significantly lower (6.2%) than that with conventional UHMWPE (11.7%) at a mean follow-up of 16 years, according to the Australian Orthopaedic Association National Joint Replacement Registry. However, HXLPE does not confer to TKA the same advantages it confers to THA. Several alternatives have been developed to prevent the release of free radicals and improve polymer mechanical properties, such as thermal treatment, phospholipid polymer 2-methacryloyloxyethyl phosphorylcholine grafting, remelting, and vitamin E addition. Among these options, vitamin E addition has reported good clinical results and wear resistance similar to that of HXLPE without vitamin E, as shown by short-term clinical studies of THA and TKA. This review aims to provide a comprehensive overview of the development and performance of UHMWPE in THA and TKA. [ABSTRACT FROM AUTHOR]

Background: The spontaneous regression of osteochondromas is rare, and only a few cases have been reported. Furthermore, the precise mechanism underlying spontaneous regression is unknown. This study aimed to examine the radiological findings of osteochondromas that had spontaneous regression and to identify potential indicators of this uncommon phenomenon in skeletally immature patients with osteochondromas. Methods: We included 28 patients (15 males and 13 females) who met the eligibility criteria between 2002 and 2019. The mean age at initial diagnosis was 9.7 years old (2–16 years). The mean follow-up period was 6.4 years (3–16 years). Results: Of the 28 patients, 10 (35.7%) had osteochondroma resolution. The osteochondroma resolved in one patient and regressed in nine. Tumor shrinkage is related to the thickness of the cartilage cap. The thickness of the cartilage cap did not correlate with age. Conclusions: Tumor shrinkage is associated with a thinner cartilage cap on magnetic resonance imaging. The thickness of the cartilage cap may be an important predictor of spontaneous regression in pediatric patients with osteochondroma. [ABSTRACT FROM AUTHOR]

Simple Summary: We hypothesized that lymphocyte to C-reactive protein ratio (LCR) could predict survival in 132 patients with soft tissue sarcoma (STS). This study aimed to determine whether LCR before treatment can predict disease-specific survival (DSS) and event-free survival (EFS). The 5-year DSS in patients with higher and lower LCR was 86.5% and 52.8%, respectively (p < 0.001). Patients with lower LCR had worse survival than those with higher LCR. The 5-year EFS in patients with higher and lower LCR was 66.2% and 31.2%, respectively (p < 0.001). On Receiver operating characteristic analysis, however, there was no significant difference in the area under curve (AUC) between CRP level (AUC = 0.72) and LCR (AUC = 0.711). LCR was found to be a poor prognostic factor for oncological outcomes using multivariate analysis. although ROC analysis could not show the superiority of LCR to CRP for predicting oncological outcomes in patients with STS. Background: Recently, the lymphocyte-to-CRP ratio (LCR) was found to have a prognostic role in many cancers. However, the clinical significance of LCR in patients with soft tissue sarcoma (STS) has not yet been established. This study aimed to determine whether LCR can predict disease-specific survival (DSS) and event-free survival (EFS) in patients with STS. Methods: In this study, 132 patients were enrolled. The mean follow-up periods were 76.5 months. Blood examinations were performed prior to treatment for all patients. Results: The 5-year DSS in patients with higher and lower LCR was 86.5% and 52.8%, respectively (p < 0.001). Patients with lower LCR had worse survival than those with higher LCR. The 5-year EFS in patients with higher and lower LCR was 66.2% and 31.2%, respectively (p < 0.001). On Receiver operating characteristic analysis, however, there was no significant difference in the area under curve (AUC) between CRP level (AUC = 0.72) and LCR (AUC = 0.711). Conclusions: LCR may be a prognostic factor for predicting oncological events in multivariate analysis, although ROC analysis could not show the superiority of LCR to CRP for predicting oncological outcomes in patients with STS. [ABSTRACT FROM AUTHOR]

Background: this study aimed to assess the long-term outcomes of (a minimum of 10-years) total hip arthroplasty with a metal-on-metal acetabular prosthesis. Methods: Eighty-nine primary total hip arthroplasties (82 patients) were performed using a Pinnacle modular metal-on-metal acetabular prosthesis. Clinical hip function outcomes were evaluated using the Japanese Orthopaedic Association hip score preoperatively and at the final follow-up. Radiological analysis was performed at the final follow-up and magnetic resonance imaging in all hips postoperatively. Results: Out of 82 patients, 17 were excluded who were followed up for <10 years. Of the remaining 65 patients (70 hips), 19 (20 hips) developed pseudotumors during 2–10 years postoperatively. After 10 and 13 years, the survival rates of revision endpoint were 93.6% and 90.4%, respectively. Clinical hip function outcomes had improved significantly at the final follow-up. In the radiological analysis, the mean cup angle of inclination and mean ratio of femoral offset on the operated hip to the contralateral hip was highest in patients with revision surgery for adverse reactions to metal debris. Conclusions: This study showed a 29.0% prevalence of pseudotumors. Some cases required revisions even after 10 years following surgery. Regular clinical surveillance is recommended for the early detection of adverse reactions to metal debris. [ABSTRACT FROM AUTHOR]

The involvement of the subtalar joint is uncommon in the early stages of rheumatoid arthritis (RA). We report a case of a 47-year-old female who had RA with isolated subtalar joint arthritis. The clinical history, magnetic resonance imaging, and pathological findings of the patient are presented. A careful evaluation of the patients for chronic ankle-to-heel pain should be conducted, and concomitant evaluation for inflammatory arthritis, including RA, should be considered. [ABSTRACT FROM AUTHOR]

Background and Objectives: Clinical studies of platelet-rich plasma (PRP) for the treatment of low back pain (LBP) have been reported; however, less is known about its long-term efficiency. Materials and Methods: This study was a long-term follow-up of a previous prospective clinical feasibility study for the use of PRP releasate (PRPr) to treat discogenic LBP patients. Among 14 patients, 11 patients were evaluated for a long-term survey. The efficacy was assessed by a visual analogue scale (VAS) for LBP intensity and the Roland-Morris Disability Questionnaire (RDQ) for LBP-related disability. Radiographic disc height was evaluated for seven patients. Results: Improvements in VAS and RDQ were sustained at an average of 5.9 years after the intradiscal injection of PRPr (p < 0.01 vs. baseline, respectively). Clinically meaningful improvements (more than 30% decrease from baseline) in VAS and RDQ were identified in 91% of patients at final survey. The radiographic measurement of disc height of PRPr-injected discs showed a mild decrease (13.8% decrease compared to baseline) during the average 5.9 years. Conclusions: The results of this study with a small number of patients suggest that the intradiscal injection of PRPr has a safe and efficacious effect on LBP improvement for more than 5 years after treatment. Further large-scale studies would be needed to confirm the clinical evidence for the use of PRPr for the treatment of patients with discogenic LBP. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are small non-coding RNAs that play a mostly post-transcriptional regulatory role in gene expression. Using RAW264.7 pre-osteoclast cells and genome-wide expression analysis, we identified a set of miRNAs that are involved in osteoclastogenesis. Based on in silico analysis, we specifically focused on miR-222-3p and evaluated its role in osteoclastogenesis. The results show that the inhibitor of miR-222-3p upregulated the mRNA levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) and tartrate-resistant acid phosphatase (TRAP), while its mimicking agent downregulated their mRNA levels. Western blot analysis showed that its inhibitor increased the protein levels of TRAP and cathepsin K, while its mimicking agent decreased their levels. Genome-wide mRNA expression analysis in the presence and absence of receptor activator of nuclear factor κ-B ligand (RANKL) predicted c-Src as a potential regulatory target of miR-222-3p. Live cell imaging using a fluorescence resonance energy transfer (FRET) technique revealed that miR-222-3p acted as an inhibitor of c-Src activity, and a partial silencing of c-Src suppressed RANKL-induced expression of TRAP and cathepsin K, as well as the number of multi-nucleated osteoclasts and their pit formation. Collectively, the study herein demonstrates that miR-222-3p serves as an inhibitor of osteoclastogenesis and c-Src mediates its inhibition of cathepsin K and TRAP. [ABSTRACT FROM AUTHOR]

Simple Summary: We aimed to elucidate the long-term outcomes of a distal femur reconstruction system in 125 patients with bone and soft tissue tumors. Implant survival rates at 10 and 15 years were 58.5% and 39.4%. Stem breakage should be considered in patients with cementless and/or smaller femoral stem sizes. Aseptic loosening should be considered in patients with a cement system after 10 years. Background: The distal femur is a common site of bone tumors. After surgical resection, prosthetic replacement is a major reconstruction method. We aimed to elucidate the long-term outcomes of the Kyocera Modular Limb Salvage (KMLS) systems after resection of tumors in the distal part of the femur. Methods: Between 1998 and 2014, 125 patients were treated at 14 institutions. There were 59 males and 66 females, with a mean age of 35 years. The mean follow-up period was 132 months. Results: There had been 65 additional surgeries, including 56 revisions and 9 amputations: 15 for aseptic loosening, 14 for stem breakage, 13 for deep infection, 13 for rotator-hinge bushing failure, 5 for local recurrence, and 5 for others. Implant survival rates at 10 and 15 years were 58.5% and 39.4%. The cumulative incidence of 15-year revision for femoral stem breakage was 31.7% in patients with cementless fixation. The 15-year cumulative incidence of revision for aseptic loosening was 19.8% in patients with cement fixation. Conclusions: KMLS systems represent a reliable system with long-term results. Stem breakage should be considered in patients with cementless and/or smaller femoral stem sizes. Aseptic loosening should be considered in patients with cement systems after 10 years. [ABSTRACT FROM AUTHOR]

Ti–6Al–4V alloy has been widely used in the orthopedic and dental fields owing to its high mechanical strength and biocompatibility. However, this alloy has a poor bone-bonding capacity, and its implantation often causes loosening. Osteoporosis increases with the aging of the population, and bisphosphonate drugs such as alendronate and minodronate (MA) are used for the medical treatment. Reliable and multifunctional implants showing both bone bonding and drug releasing functions are desired. In this study, we developed a novel organic-inorganic composite layer consisting of MA-containing gelatin and calcium-deficient calcium titanate (cd–CT) with high bone-bonding and scratch resistance on Ti–6Al–4V alloy. The alloy with the composite layer formed apatite within 7 days in a simulated body fluid and exhibited high scratch resistance of an approximately 50 mN, attributable to interlocking with cd ± CT. Although the gelatin layer almost completely dissolved in phosphate-buffered saline within 6 h, its dissolution rate was significantly suppressed by a subsequent thermal crosslinking treatment. The released MA was estimated at more than 0.10 μmol/L after 7 days. It is expected that the Ti alloy with the MA-containing gelatin and cd–CT composite layer will be useful for the treatment of osteoporosis bone. [ABSTRACT FROM AUTHOR]

Clinical application of platelet-rich plasma is gaining popularity in treating low back pain (LBP). This study investigated the efficacy and safety of platelet-rich plasma releasate (PRPr) injection into degenerated discs of patients with discogenic LBP. A randomized, double-blind, active-controlled clinical trial was conducted. Sixteen patients with discogenic LBP received an intradiscal injection of either autologous PRPr or corticosteroid (CS). Patients in both groups who wished to have PRPr treatment received an optional injection of PRPr eight weeks later. The primary outcome was change in VAS from baseline at eight weeks. Secondary outcomes were pain, disability, quality of life (QOL), image analyses of disc degeneration, and safety for up to 60 weeks. The VAS change at eight weeks did not significantly differ between the two groups. Fifteen patients received the optional injection. Compared to the CS group, the PRPr group had a significantly improved disability score at 26 weeks and walking ability scores at four and eight weeks. Radiographic disc height and MRI grading score were unchanged from baseline. PRPr caused no clinically important adverse events. PRPr injection showed clinically significant improvements in LBP intensity equal to that of CS. PRPr treatment relieved pain, and improved disability and QOL during 60 weeks of observation. [ABSTRACT FROM AUTHOR]

Simple Summary: Breast cancer is a common malignancy in women in the US. While brain metastases frequently occur from advanced breast cancer, current treatments are of limited effectiveness. In this study, we found that osteocytes and their conditioned medium (CM) presented the tumor-suppressing capability, and the overexpression of Lrp5, IL1ra, and β-catenin enhanced their anti-tumor actions. In a mouse model, the administration of osteocytes and their CM inhibited the progression of mammary tumors and tumors in the bone and brain. The minimally invasive administration allowed tumor-suppressing factors in CM to diffuse into the brain. Besides p53 and Trail, mass spectrometry-based whole-genome proteomics revealed that extracellular histone H4 was enriched in CM and acted as a tumor suppressor. We also observed that Lrp5-overexpressing mesenchymal stem cells presented the tumor-suppressing capability. The brain is a common site of metastasis from advanced breast cancer but few effective treatments are available. We examined a therapeutic option with a conditioned medium (CM), focusing on the role of Lrp5 and β-catenin in Wnt signaling, and IL1ra in osteocytes. Osteocytes presented the innate anti-tumor effect and the overexpression of the above genes strengthened their action. In a mouse model, the injection of their CM inhibited mammary tumors and tumor-driven osteolysis. Importantly, Lrp5- and/or IL1ra-overexpressing osteocytes or the local administration of β-catenin-overexpressing CM markedly inhibited brain tumors. In the transport analysis, tumor-suppressing factors in CM were shown to diffuse through the skull. Mechanistically, the CM with overexpression of the above genes downregulated oncogenic genes such as MMP9, Runx2, TGFβ, and Snail in breast cancer cells. Also, the CM with β-catenin overexpression downregulated CXCL1 and CXCL5 and upregulated tumor suppressors such as LIMA1, DSP, p53, and TRAIL in breast cancer cells. Notably, whole-genome proteomics revealed that histone H4 was enriched in CM and acted as an atypical tumor suppressor. Lrp5-overexpressing MSCs were also shown to act as anti-tumor agents. Collectively, this study demonstrated the therapeutic role of engineered CM in brain tumors and the tumor-suppressing action of extracellular histone H4. The result sheds light on the potential CM-based therapy for breast cancer-associated brain metastases in a minimally invasive manner. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate skeletal pain associated with osteoporosis and to examine the inhibitory effects of cytotoxic T lymphocyte-associated antigen-4Ig (CTLA-4Ig) administration in ovariectomized (OVX) mice. Eight-week-old female ddY mice were assigned to three groups: sham-operated mice (SHAM) treated with vehicle, OVX mice treated with vehicle (OVX), and OVX mice treated with CTLA-4Ig (CTLA-4Ig). Vehicle or CTLA-4Ig was injected intraperitoneally, starting immediately after surgery. After 4 weeks of treatment, mechanical sensitivity was examined, and the bilateral hind limbs were removed and evaluated by micro-computed tomography, immunohistochemical analyses, and messenger RNA expression analysis. Ovariectomy induced bone loss and mechanical hyperalgesia in the hindlimbs. CTLA-4Ig treatment prevented bone loss in the hindlimbs compared to vehicle administration in the OVX group. Moreover, mechanical hyperalgesia was significantly decreased in the CTLA-4Ig treatment group in comparison to the OVX group. The expression levels of tumor necrosis factor-α (TNF-α) and sclerostin (SOST), as well as the number of osteoclasts, were increased, and the expression level of Wnt-10b was decreased in the OVX group compared with the SHAM group, whereas these parameters were improved in the CTLA-4Ig group compared with the OVX group. The novelty of this research is that CTLA-4Ig administration prevented bone loss and mechanical hyperalgesia induced by ovariectomy in the hindlimbs. [ABSTRACT FROM AUTHOR]

Interleukin-6 (IL-6) is widely accepted to stimulate osteoclasts. Our aim in this study was to examine whether the inhibitory effect of IL-6 on bone loss and skeletal pain associated with osteoporosis in hindlimb-unloaded (HU) mice in comparison with bisphosphonate. Eight-week-old male ddY mice were tail suspended for 2 weeks. Starting immediately after reload, vehicle (HU group), alendronate (HU-ALN group), or anti-IL-6 receptor antibody (HU-IL-6i group) was injected subcutaneously. After a 2-week treatment, pain-related behavior was examined using von Frey filaments. The bilateral distal femoral and proximal tibial metaphyses were analyzed three-dimensionally with micro-computed tomography. Calcitonin gene-related peptide (CGRP) expressions in dorsal root ganglion (DRG) neurons innervating the hindlimbs were examined using immunohistochemistry. HU mice with tail suspension developed bone loss. The HU mice showed mechanical hyperalgesia in the hindlimbs and increased CGRP immunoreactive neurons in the L3-5 DRG. Treatment with IL-6i and ALN prevented HU-induced mechanical hyperalgesia and upregulation of CGRP expressions in DRG neurons. Furthermore, ALN but not IL-6i prevented HU-induced bone loss. In summary, treatment with IL-6i prevented mechanical hyperalgesia in hindlimbs and suppressed CGRP expressions in DRG neurons of osteoporotic models. The novelty of this research suggests that IL-6 is one of the causes of immobility-induced osteoporotic pain regardless improvement of bone loss. [ABSTRACT FROM AUTHOR]

To assess the long-term effects of tadalafil, a therapeutic agent for fetal growth restriction (FGR), we evaluated the developmental progress of 1.5-year-old infants whose mothers had taken tadalafil during pregnancy. Twenty-four infants were assessed. We evaluated infant body weight, height, and head circumference, and performed the Kyoto Scale of Psychological Development (KSPD) test, a standardized developmental assessment covering Postural–Motor (P–M), Cognitive–Adaptive (C–A), and Language-Social (L–S) functions. The sum score was converted to a developmental quotient (DQ). The mean gestational week of the included cases was 36.1 (29–39) weeks, and the mean birth weight was 1841 (874–2646) g. Twenty-one and 20 out of the 24 cases, respectively, attained body weight and height similar to those of age-matched normal infants (within the 3rd percentile); all cases caught up in head circumference. KSPD was performed for 18 cases at 1.5 years of corrected age. The mean DQ scores were 87 (in total): 82 in P–M, 90 in C–A, and 88 in L–S. The total DQ score in one case (5.6%) was less than 70, and ranged from 70 to 85 in five cases (27.7%), and was more than 85 in 11 cases (61.1%). The growth and development of infants born of tadalafil-treated mothers seem to show good progress at a corrected age of 1.5 years. [ABSTRACT FROM AUTHOR]

Interleukin-6 (IL-6) affects the key parameters of oncogenesis, which increases the cell resistance to apoptosis, the proliferation of cancer cells, angiogenesis, invasion, malignancy, and the ability of tumor cells to respond to anticancer therapy. This study aimed to elucidate the association between IL-6 and IL-6 receptor (IL-6R) expression in tissues and clinical outcomes in patients with soft tissue sarcomas (STSs) because, to our knowledge, this has not been done before. We enrolled 86 patients with histologically-proven localized STSs who underwent surgical resection. The cohort included 48 men and 38 women, with a mean age of 65.6 years. The mean follow-up duration was 40.5 months. The expression of IL-6 and IL-6R was immunohistochemically determined. We analyzed prognostic factors for overall survival (OS) and metastasis-free survival (MFS). High IL-6 expression was observed in 23.3% (20/86), high IL-6R expression in 44.2% (38/86), and high expression of both in 16.3% (14/86) of patients. Multivariate analysis showed that a high expression of both IL-6 and IL-6R was a prognostic factor for OS and MFS. We found that this high expression indicated that the patient had a poor prognosis for OS and MFS. [ABSTRACT FROM AUTHOR]

Aim: Primary malignant bone tumor osteosarcoma can metastasize to the lung. Diminishing lung metastasis would positively affect the prognosis of patients. Our previous studies demonstrated that highly metastatic osteosarcoma cell lines are significantly softer than low-metastasis cell lines. We therefore hypothesized that increasing cell stiffness would suppress metastasis by reducing cell motility. In this study, we tested whether carbenoxolone (CBX) increases the stiffness of LM8 osteosarcoma cells and prevents lung metastasis in vivo., Methods: We evaluated the actin cytoskeletal structure and polymerization of CBX-treated LM8 cells using actin staining. Cell stiffness was measured using atomic force microscopy. Metastasis-related cell functions were analyzed using cell proliferation, wound healing, invasion, and cell adhesion assays. Furthermore, lung metastasis was examined in LM8-bearing mice administered with CBX., Results: Treatment with CBX significantly increased actin staining intensity and stiffness of LM8 cells compared with vehicle-treated LM8 cells ( p < 0.01). In Young's modulus images, compared with the control group, rigid fibrillate structures were observed in the CBX treatment group. CBX suppressed cell migration, invasion, and adhesion but not cell proliferation. The number of LM8 lung metastases were significantly reduced in the CBX administration group compared with the control group ( p < 0.01)., Conclusion: In this study, we demonstrated that CBX increases tumor cell stiffness and significantly reduces lung metastasis. Our study is the first to provide evidence that reducing cell motility by increasing cell stiffness might be effective as a novel anti-metastasis approach in vivo.

Ewing sarcoma is an aggressive and the second most common bone tumor in adolescent and young adult patients. The 5-year survival rate is 60-70% for localized disease but 30% for patients with metastases. Here, we aimed to identify a therapeutic target for Ewing sarcoma and evaluate antibody-based therapeutic agents using in vitro and in vivo models. We identified G protein-coupled receptor 64 (GPR64) as a therapeutic target for Ewing sarcoma via next-generation RNA-sequencing. GPR64v205 mRNA was expressed in HTB166, A673, MG63, 143B, HS-Sy II, and HT1080 cell lines as well as in Ewing sarcoma, undifferentiated pleomorphic sarcoma, leiomyosarcoma, dedifferentiated liposarcoma, and synovial sarcoma tissues. GPR64 expression was observed in 62.5% of sarcoma cases and was overexpressed in 33.9% cases. GPR64-specific monoclonal antibodies were tested as near-infrared probes for in vivo imaging using subcutaneous tumor mouse xenografts. Fluorescence intensity was stronger for the AF700-labeled anti-GPR64 antibody than that for the AF700-labeled isotype control antibody. GPR64 was detected in engrafted tumors of A673, 143B, HT1080, and the epididymis but not in other resected tissues. The anti-GPR64 antibody showed excellent binding to GPR64-positive tumors but not to healthy tissues. This antibody has potential for drug delivery in the antibody-based treatment of sarcomas.

Osteosarcoma is the most common primary malignant bone tumor. The cause of death due to osteosarcoma is typically a consequence of metastasis to the lung. Controlling metastasis leads to improved prognosis for osteosarcoma patients. The cell stiffness of several tumor types is involved in metastatic potential; however, it is unclear whether the metastatic potential of osteosarcoma depends on cell stiffness. In this study, we analyzed the cell stiffness of the low metastatic Dunn cell line and its highly metastatic LM8 subline, and compared actin organization, cell proliferation, and metastasis. Actin cytoskeleton, polymerization, stiffness, and other cellular properties were analyzed. The organization of the actin cytoskeleton was evaluated by staining F-actin with Alexa Fluor 488 phalloidin. Cell stiffness was measured using Atomic Force Microscopy (AFM). Cell proliferation, migration, invasion, and adhesion were also evaluated. All experiments were performed using mouse osteosarcoma cell lines cultured in the absence and presence of cytochalasin. In LM8 cells, actin polymerization was strongly suppressed and actin levels were significantly lower than in Dunn cells. Stiffness evaluation revealed that LM8 cells were significantly softer than Dunn. Young's modulus images showed more rigid fibrillar structures were present in Dunn cells than in LM8 cells. LM8 cells also exhibited a significantly higher proliferation. The migration and invasion potential were also higher in LM8 cells, whereas the adhesion potential was higher in Dunn cells. The administration of cytochalasin resulted in actin filament fragmentation and decreased actin staining intensity and cell stiffness in both LM8 and Dunn cells. Cells with high metastatic potential exhibited lower actin levels and cell stiffness than cells with low metastatic potential. The metastatic phenotype is highly correlated to actin status and cell stiffness in osteosarcoma cells. These results suggest that evaluation of actin dynamics and cell stiffness is an important quantitative diagnostic parameter for predicting metastatic potential. We believe that these parameters represent new reliable quantitative indicators that can facilitate the development of new drugs against metastasis.

Integrated stress responses (ISR) may lead to cell death and tissue degeneration via eukaryotic translation initiation factor 2 α (eIF2α)-mediated signaling. Alleviating ISR by modulating eIF2α phosphorylation can reduce the symptoms associated with various diseases. Guanabenz is known to elevate the phosphorylation level of eIF2α and reduce pro-inflammatory responses. However, the mechanism of its action is not well understood. In this study, we investigated the signaling pathway through which guanabenz induces anti-inflammatory effects in immune cells, in particular macrophages. Genome-wide mRNA profiling followed by principal component analysis predicted that colony stimulating factor 2 (Csf2, or GM-CSF as granulocyte macrophage colony stimulating factor) is involved in the responses to guanabenz. A partial silencing of Csf2 or eIF2α by RNA interference revealed that Interleukin-6 (IL6), Csf2, and Cyclooxygenase-2 (Cox2) are downregulated by guanabenz-driven phosphorylation of eIF2α. Although expression of IL1β and Tumor Necrosis Factor-α (TNFα) was suppressed by guanabenz, their downregulation was not directly mediated by eIF2α signaling. Collectively, the result herein indicates that anti-inflammatory effects by guanabenz are mediated by not only eIF2α-dependent but also eIF2α-independent signaling.