A novel mechanism behind irreversible development of cartilage degradation driven articular cartilage defects revealed by rat model: The chain reaction initiated by extracellular vesicles delivered LOC102546541.

BACKGROUND: Articular cartilage defects (ACD) are injuries with a diameter greater than 3 mm, resulting from wear and tear on joints. When the diameter of the defect exceeds 6 mm, it can further damage the surrounding joint cartilage, causing osteoarthritis (OA). Try to explain why OA is an irreversible disease, we hypothesize that damaged articular chondrocytes (DAC) may have reduced capacities to repair cartilage because its extracellular vesicle (EVs) that might directly contribute to OA formation. METHODS: In this study, DAC-EVs and AC-EVs were isolated using ultracentrifugation. Next-generation sequencing was employed to screen for a pathogenic long non-coding RNA (lncRNA). After verifying its function in vitro, the corresponding small interfering RNA (siRNA) was constructed and loaded into extracellular vesicles, which were then injected into the knee joint cavities of rats. RESULTS: The results revealed that DAC-EVs packaged lncRNA LOC102546541 acts as a competitive endogenous RNA (ceRNA) of MMP13, down-regulating miR-632. Consequently, the function of MMP13 in degrading the extracellular matrix is enhanced, promoting the development of osteoarthritis. CONCLUSIONS: This study uncovered a novel mode of OA pathogenesis using rat models, which DAC deliver pathogenic LOC102546541 packaged EVs to normal articular chondrocytes, amplifying the degradation of the extracellular matrix. Nonetheless, the functions of highly homologous human gene of LOC102546541 need to be verified in the future.

Proteome-wide Mendelian randomization identifies therapeutic targets for ankylosing spondylitis.

Background: Ankylosing Spondylitis (AS) is a chronic inflammatory disorder which can lead to considerable pain and disability. Mendelian randomization (MR) has been extensively applied for repurposing licensed drugs and uncovering new therapeutic targets. Our objective is to pinpoint innovative therapeutic protein targets for AS and assess the potential adverse effects of druggable proteins. Methods: We conducted a comprehensive proteome-wide MR study to assess the causal relationships between plasma proteins and the risk of AS. The plasma proteins were sourced from the UK Biobank Pharma Proteomics Project (UKB-PPP) database, encompassing GWAS data for 2,940 plasma proteins. Additionally, GWAS data for AS were extracted from the R9 version of the Finnish database, including 2,860 patients and 270,964 controls. The colocalization analysis was executed to identify shared causal variants between plasma proteins and AS. Finally, we examined the potential adverse effects of druggable proteins for AS therapy by conducting a phenome-wide association study (PheWAS) utilizing the extensive Finnish database in version R9, encompassing 2,272 phenotypes categorized into 46 groups. Results: The findings revealed a positive genetic association between the predicted plasma levels of six proteins and an elevated risk of AS, while two proteins exhibited an inverse association with AS risk (P fdr < 0.05). Among these eight plasma proteins, colocalization analysis identified AIF1, TNF, FKBPL, AGER, ALDH5A1, and ACOT13 as shared variation with AS(PPH3+PPH4>0.8), suggesting that they represent potential direct targets for AS intervention. Further phenotype-wide association studies have shown some potential side effects of these six targets (P fdr < 0.05). Conclusion: Our investigation examined the causal connections between six plasma proteins and AS, providing a comprehensive understanding of potential therapeutic targets.

The rat as a novel model for chronic rotator cuff injuries.

Chronic rotator cuff injuries (CRCIs) still present a great challenge for orthopaedics surgeons. Many new therapeutic strategies are developed to facilitate repair and improve the healing process. However, there is no reliable animal model for chronic rotator cuff injury research. To present a new valuable rat model for future chronic rotator cuff injuries (CRCIs) repair studies, and describe the changes of CRCIs on the perspectives of histology, behavior and MRI. Sixty male Wistar rats were enrolled and underwent surgery of the left shoulder joint for persistent subacromial impingement. They were randomly divided into experimental group (n = 30, a 3D printed PEEK implant shuttled into the lower surface of the acromion) and sham operation group (n = 30, insert the same implant, but remove it immediately). Analyses of histology, behavior, MRI and inflammatory pain-related genes expression profiles were performed to evaluate the changes of CRCIs. After 2-weeks running, the rats in the experimental group exhibited compensatory gait patterns to protect the injured forelimb from loading after 2-weeks running. After 8-weeks running, the rats in the experimental group showed obvious CRCIs pathological changes: (1) acromion bone hyperplasia and thickening of the cortical bone; (2) supraspinatus muscle tendon of the humeral head: the bursal-side tendon was torn and layered with disordered structure, forming obvious gaps; the humeral-side tendon is partially broken, and has a neatly arranged collagen. Partial fat infiltration is found. The coronal T2-weighted images showed that abnormal tendon-to-bone junctions of the supraspinatus tendon. The signal intensity and continuity were destroyed with contracted tendon. At the nighttime, compared with the sham operation group, the expression level of IL-1ß and COX-2 increased significantly (P = 0063, 0.0005) in the experimental group. The expression of COX-2 in experimental group is up-regulated about 1.5 times than that of daytime (P = 0.0011), but the expression of IL-1ß, TNF-a, and NGF are all down-regulated (P = 0.0146, 0.0232, 0.0161). This novel rat model of chronic rotator cuff injuries has the similar characteristics with that of human shoulders. And it supplies a cost-effective, reliable animal model for advanced tissue engineered strategies and future therapeutic strategies.

A novel fluffy PLGA/HA composite scaffold for bone defect repair.

Treatment of bone defects remains crucial challenge for successful bone healing, which arouses great interests in designing and fabricating ideal biomaterials. In this regard, the present study focuses on developing a novel fluffy scaffold of poly Lactide-co-glycolide (PLGA) composites with hydroxyapatite (HA) scaffold used in bone defect repair in rabbits. This fluffy PLGA/HA composite scaffold was fabricated by using multi-electro-spinning combined with biomineralization technology. In vitro analysis of human bone marrow mesenchymal stem cells (BMSCs) seeded onto fluffy PLGA/HA composite scaffold showed their ability to adhere, proliferate and cell viability. Transplant of fluffy PLGA/HA composite scaffold in a rabbit model showed a significant increase in mineralized tissue production compared to conventional and fluffy PLGA/HA composite scaffold. These findings are promising for fluffy PLGA/HA composite scaffolds used in bone defects.

Exosomes derived from mesenchymal stem cells rescue cartilage injury in osteoarthritis through Ferroptosis by GOT1/CCR2 expression.

Osteoarthritis is a prevalent joint disease that significantly affects the daily life of the elderly and is one of the primary causes of disability in this population. This study aims to evaluate the potential pro-inflammatory effects and molecular mechanism of Mesenchymal stem cells-derived exosomes (MSC-Exos) in Osteoarthritis. Bilateral ovariectomy was carried out to induce osteoporosis under anesthesia for the mice. MC3T3-E1 cells were induced for 14 days.HE staining, Safranin O staining and Biomechanical parameter analysis were used in this experiment. MSC-Exos improved osteoarthritis in a mouse model by reducing inflammation levels, preventing ferroptosis, and inducing expression of GOT1/CCR2 to regulate ferroptosis. MSC-Exos also promoted cell growth and osteogenic differentiation of bone cells in an in vitro model. Inhibition of GOT1 reduced the effects of MSC-Exos on cell growth and osteogenic differentiation in an osteoarthritis model. MSC-Exos induce Nrf2/HO-1 expression through the GOT1/CCR2 signaling pathway, resulting in the reduction of Ferroptosis. However, inhibition of Nrf2 reduces the effectiveness of MSC-Exos in treating Osteoarthritis.The results of this study suggest that the GOT1/CCR2/Nrf2/HO-1 signaling pathway plays a crucial role in MSC-Exos-mediated reduction of Ferroptosis in macrophages during Osteoarthritis. These findings may provide a potential therapeutic approach for Osteoarthritis and other orthopedic conditions.

All-inside technique versus in situ transtendon repair for Ellman â ¢ partial articular supraspinatus tendon avulsion.

BACKGROUND: The current surgical techniques for repairing Ellman Ⅲ partial articular supraspinatus tendon avulsion (PASTA) is mainly tear completion followed by repair and in situ transtendon repair, and both techniques have been proven to have good clinical effects. In situ transtendon repair is more widely used because of its high performance in preserving the integrity of the bursal side supraspinatus tendon. However, there is still some scope for improvement. Our purpose was to compare the clinical outcomes of the all-inside repair technique and in situ transtendon repair for Ellman III PASTA. METHODS: A retrospective comparative study was conducted on 56 patients who suffered from Ellman Ⅲ PASTA and underwent rotator cuff repair; 28 patients were treated with the all-inside technique (group A), and 28 patients were treated with the transtendinous technique (group B). All patients were followed up for at least 2 years. The visual analog scale (VAS), Constant, and American Shoulder and Elbow Surgeons (ASES) scores were used to evaluate the patient's shoulder joint function before surgery, 1 month and 3 months after surgery, and at the last time of follow-up. RESULTS: Group A showed superiority in pain and functional improvement 1 month after the operation: the VAS score, 2.8 ± 0.3 in group A vs. 4.7 ± 0.4 in group B (P = .042); Constant score, 73 ± 5 in group A vs. 60 ± 6 in group B (P = .038); and ASES score, 75 ± 5 in group A vs. 58 ± 7 in group B (P = .043), whereas there was no statistical difference in group B. However, 3 months after surgery and at the last follow-up, the VAS, Constant, and ASES scores in both groups were significantly improved (P < .01), and there was no significant statistical difference between the groups (P > .05). Magnetic resonance imaging showed that the repaired rotator cuff tendon-bone healed well; at the last follow-up, all patients were in good function, the pain was completely relieved, and no revision was performed in both groups. CONCLUSION: Arthroscopic repair of Ellman Ⅲ PASTA provided satisfactory functional improvements and pain relief regardless of the all-inside and in situ transtendon repair techniques. However, the all-inside repair technique is more beneficial due to its dual function in preserving the intact bursa and avoidance of uneven tension of the articular side, which is advantageous to early postoperative rehabilitation.

Biofeedback electrostimulation for bionic and long-lasting neural modulation.

Invasive electrical stimulation (iES) is prone to cause neural stimulus-inertia owing to its excessive accumulation of exogenous charges, thereby resulting in many side effects and even failure of nerve regeneration and functional recovery. Here, a wearable neural iES system is well designed and built for bionic and long-lasting neural modulation. It can automatically yield biomimetic pulsed electrical signals under the driven of respiratory motion. These electrical signals are full of unique physiological synchronization can give biofeedback to respiratory behaviors, self-adjusting with different physiological states of the living body, and thus realizing a dynamic and biological self-matched modulation of voltage-gated calcium channels on the cell membrane. Abundant cellular and animal experimental evidence confirm an effective elimination of neural stimulus-inertia by these bioelectrical signals. An unprecedented nerve regeneration and motor functional reconstruction are achieved in long-segmental peripheral nerve defects, which is equal to the gold standard of nerve repair -- autograft. The wearable neural iES system provides an advanced platform to overcome the common neural stimulus-inertia and gives a broad avenue for personalized iES therapy of nerve injury and neurodegenerative diseases.

Pathologic mechanism of hidden blood loss after total knee arthroplasty: oxidative stress induced by free fatty acids.

BACKGROUND: Hidden blood loss (HBL) often occurs after joint replacement; however, the mechanism of HBL is not clear. We conducted a prospective study to analyze the correlation between high-level free fatty acids (FFA) and erythrocyte injury, and explore the pathologic mechanism of hidden blood loss (HBL) after total knee arthroplasty (TKA). METHODS: Perioperative blood indexes were tested in 120 patients who underwent unilateral total knee replacement for end-stage knee osteoarthritis. The changes in FFA, reactive oxygen species (ROS), hemoglobin (Hb), and red blood cells (RBC) in the blood samples were detected. The activity of glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD) and hydrogen peroxide (H2O2) levels were measured. Morphologic changes of blood cells were analyzed under a microscope. RESULTS: HBL occurred in all patients after TKA. The Hb and RBC decreased significantly 24 h after surgery (P <0.05), while FFA and ROS concentration were substantially elevated, and heteromorphic red blood cells appeared under the microscope. The hemoglobin content decreased to its lowest level at 48 h after the operation (P<0.01). With the increase of FFA and ROS levels, HBL appeared more obvious (P<0.01). GSH-PX activity, T-SOD activity, and H2O2 levels significantly decreased compared to preoperative tested samples (P<0.01). Microscopically, atypical erythrocytes increased significantly with cellular rupture and lysis identified. CONCLUSIONS: High levels of FFA in blood can induce oxidative stress and damage red blood cells, leading to the occurrence of HBL after surgery. TRIAL REGISTRATION: Chinese Clinical Trials Registry (the trial number: ChiCTR17010681, URL: http://www.chictr.org.cn).

Physiologically Self-Regulated, Fully Implantable, Battery-Free System for Peripheral Nerve Restoration.

The long-segment peripheral nerve injury (PNI) represents a global medical challenge, leading to incomplete nerve tissue recovery and unsatisfactory functional reconstruction. However, the current electrical stimulation (ES) apparatuses fail perfect nerve repair due to their inability of the variable synchronous self-regulated function with physiological states. It is urgent to develop an implantable ES platform with physiologically adaptive function to provide instantaneous and nerve-preferred ES. Here, a physiologically self-regulated electrical signal is generated by integrating a novel tribo/piezoelectric hybrid nanogenerator with a nanoporous nerve guide conduit to construct a fully implantable neural electrical stimulation (FI-NES) system. The optimal neural ES parameters completely originate from the body itself and are highly self-responsive to different physiological states. The morphological evaluation, representative protein expression level, and functional reconstruction of the regenerated nerves are conducted to assess the PNI recovery process. Evidence shows that the recovery effect of 15 mm length nerve defects under the guidance of the FI-NES system is significantly close to the autograft. The designed FI-NES system provides an effective method for long-term accelerating the recovery of PNI in vivo and is also appropriate for other tissue injury or neurodegenerative diseases.

Proteomic analysis reveals rotator cuff injury caused by oxidative stress.

BACKGROUND AND AIMS: Rotator cuff tendinopathy is common and is related to pain and dysfunction. However, the pathological mechanism of rotator cuff injury and shoulder pain is unclear. Objective: to investigate the pathological mechanism of rotator cuff injury and shoulder pain, and screen out the marker proteins related to rotator cuff injury by proteomics. METHODS: Subacromial synovium specimens were collected from patients undergoing shoulder arthroscopic surgery. The experimental group were patients with rotator cuff repair surgery, and the control group were patients with habitual dislocation of the shoulder joint. Pathological examination was performed, and then followed by non-labeled quantitative proteomic detection. Finally, from analysis of the biological information of the samples, specific proteins related to rotator cuff injury and shoulder pain were deduced by functional analysis of differential proteins. RESULTS: All the patients in experimental groups were representative. A large number of adipocytes and inflammatory cells were found in the pathological sections of the experimental group; the proteomics analysis screen identified 80 proteins with significant differences, and the analysis of protein function revealed that S100A11 (p = 0.011), PLIN4 (p = 0.017), HYOU1 (p = 0.002) and CLIC1 (p = 0.007) were closely related to oxidative stress and chronic inflammation. CONCLUSION: Rotator cuff injury is closely related to oxidative stress and chronic inflammatory response, and the results suggest that the expression of S100A11, PLIN4, HYOU1 and CLIC1 in the synovium of rotator cuff injury provides a new marker for the study of its pathological mechanism.