Articular Cartilage Degeneration Research Articles

Evaluation of the Efficacy of Cryopreserved Human Umbilical Cord Tissue Allografts to Augment Functional and Pain Outcome Measures in Patients with Knee Osteoarthritis: An Observational Data Collection Study

The primary objective of this study is to report the initial efficacy data observed with the use of cryopreserved human umbilical tissue allografts for supplementation in patients with symptomatic degeneration of load-bearing articular cartilage in the knee joint. Our primary endpoints were pain, stiffness, and functional recovery scores. In this ongoing study, 55 participants (age 56–93 years) received a single Wharton’s jelly tissue allograft application. The study dose consisted of 150 mg of Wharton’s jelly allograft suspended in approximately 2 mL of sterile sodium chloride 0.9% solution (normal saline). Each study knee application was performed under ultrasound guidance in a physician’s office. The research methodology consisted of Numeric Pain Rating Scale (NPRS) scores and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) subsection scores including pain, stiffness, and physical function. Study enrollment consisted of 55 patients followed for a post-application duration of 90 days. No adverse events or adverse reactions were reported. The results demonstrated statistically significant improvements of NPRS and WOMAC in initial versus 90-day examination. The data represent Wharton’s jelly tissue allograft applications are safe, non-surgical, and efficacious for patients with symptomatic osteoarthritis of the knee.

Risk factors of cartilage lesion after anterior cruciate ligament reconstruction.

Anterior cruciate ligament injury is the most common sports injury in orthopaedics, which can adversely affect knee joint function and exercise of patients. Using arthroscopy to reconstruct the anterior cruciate ligament has become the first choice for treating anterior cruciate ligament rupture. However, different degrees of articular cartilage injury of the knee can be observed in patients after anterior cruciate ligament reconstruction. More importantly, the articular cartilage injury after anterior cruciate ligament reconstruction indicates that it will develop into osteoarthritis in the long term. It is of great significance to fully understand the factors that lead to the occurrence and development of cartilage injury. This article reviews the effects of surgical methods, meniscus status, different grafts, time from injury to surgical intervention, postoperative knee joint stability, postoperative rehabilitation, knee joint anatomical factors, and demographic characteristics of patients on articular cartilage degeneration after anterior cruciate ligament reconstruction. The present review provides insights into the anterior cruciate ligament reconstruction, which can be used to investigate new treatment strategies to delay and prevent the progress of osteoarthritis. At the same time, it provides a holistic understanding of the influence of multiple factors on cartilage lesions after anterior cruciate ligament reconstruction.

Interferon-gamma modulates articular chondrocyte and osteoblast metabolism through protein kinase R-independent and dependent mechanisms

Osteoarthritis (OA) affects multiple tissues of the synovial joint and is characterised by articular cartilage degeneration and bone remodelling. Interferon-γ (IFN-γ) is implicated in osteoarthritis pathology exerting its biological effects via various mechanisms including activation of protein kinase R (PKR), which has been implicated in inflammation and arthritis. This study investigated whether treatment of articular cartilage chondrocytes and osteoblasts with IFN-γ could induce a degradative phenotype that was mediated through the PKR signalling pathway. IFN-γ treatment of chondrocytes increased transcription of key inflammatory mediators (TNF-α, IL-6), matrix degrading enzymes (MMP-13), the transcription factor STAT1, and PKR. Activation of PKR was involved in the regulation of TNF-α, IL-6, and STAT1. In osteoblasts, IFN-γ increased human and mouse STAT1, and human IL-6 through a mechanism involving PKR. ALP, COL1A1 (human and mouse), RUNX2 (mouse), and PHOSPHO1 (mouse) were decreased by IFN-γ. The number of PKR positive cells were increased in post-traumatic OA (PTOA). This study has revealed that IFN-γ propagates inflammatory and degenerative events in articular chondrocytes and osteoblasts via PKR activation. Since IFN-γ and PKR signalling are both activated in early PTOA, these mechanisms are likely to contribute to joint degeneration after injury and might offer attractive targets for therapeutic intervention.

Loofah-chitosan and poly (−3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) based hydrogel scaffolds for meniscus tissue engineering applications

The meniscus is a fibrocartilaginous tissue that is very important for the stability of the knee joint. However, it has a low ability to heal itself, so damage to it will always lead to articular cartilage degeneration. The goal of this study was to make a new type of meniscus scaffold made of chitosan, loofah mat, and PHBV nanofibers, as well as to describe hydrogel composite scaffolds in terms of their shape, chemical composition, mechanical properties, and temperature. Three different concentrations of genipin (0.1, 0.3, and 0.5 %) were used and the optimal crosslinker concentration was 0.3 % for Chitosan/loofah (CL) and Chitosan/loofah/PHBV fiber (CLF). Scaffolds were seeded using undifferentiated MSCs and incubated for 21 days to investigate the chondrogenic potential of hydrogel scaffolds. Cell proliferation analyses were performed using WST-1 assay, GAG content was analyzed, SEM and fluorescence imaging observed morphologies and cell attachment, and histological and immunohistochemical studies were performed. The in vitro analysis showed no cytotoxic effect and enabled cells to attach, proliferate, and migrate inside the scaffold. In conclusion, the hydrogel composite scaffold is a promising material for engineering meniscus tissue.

Inhibition of PGE2 in Subchondral Bone Attenuates Osteoarthritis.

Aberrant subchondral bone architecture is a crucial driver of the pathological progression of osteoarthritis, coupled with increased sensory innervation. The sensory PGE2/EP4 pathway is involved in the regulation of bone mass accrual by the induction of differentiation of mesenchymal stromal cells. This study aimed to clarify whether the sensory PGE2/EP4 pathway induces aberrant structural alteration of subchondral bone in osteoarthritis. Destabilization of the medial meniscus (DMM) using a mouse model was combined with three approaches: the treatment of celecoxib, capsaicin, and sensory nerve-specific prostaglandin E2 receptor 4 (EP4)-knockout mice. Cartilage degeneration, subchondral bone architecture, PGE2 levels, distribution of sensory nerves, the number of osteoprogenitors, and pain-related behavior in DMM mice were assessed. Serum and tissue PGE2 levels and subchondral bone architecture in a human sample were measured. Increased PGE2 is closely related to subchondral bone’s abnormal microstructure in humans and mice. Elevated PGE2 concentration in subchondral bone that is mainly derived from osteoblasts occurs in early-stage osteoarthritis, preceding articular cartilage degeneration in mice. The decreased PGE2 levels by the celecoxib or sensory denervation by capsaicin attenuate the aberrant alteration of subchondral bone architecture, joint degeneration, and pain. Selective EP4 receptor knockout of the sensory nerve attenuates the aberrant formation of subchondral bone and facilitates the prevention of cartilage degeneration in DMM mice. Excessive PGE2 in subchondral bone caused a pathological alteration to subchondral bone in osteoarthritis and maintaining the physiological level of PGE2 could potentially be used as an osteoarthritis treatment.

The primary research about the impact of the estrogen deficiency on NOTCH signaling pathway in osteoarthritis

Introduction: Estrogen deficiency is the main risk factor of articular cartilage degeneration, leading to postmenopausal osteoarthritis. Moreover, there were imperfect estrogen-responsive elements that did exist in the 5’-untranslated region of Notch1 and Jagged1. So, our aim was exploring the relationship between estrogen deficiency and the Notch signaling in postmenopausal osteoarthritis.
 Methods: These rats were divided into four groups as follows1. bilateral ovariectomy and intra-articular injection of collagenase, 2. bilateral ovariectomy, 3. intra-articular injection of collagenase, 4. intra-articular injection of normal saline. Histological and immunohistochemical staining of joint cartilage were used to evaluate the osteoarthritis model, including the morphological changes of articular cartilage and the relative expression of osteoarthritic biomarkers (IL-1β, MMP-1, and type II collagen). The Notch signaling expression (Notch2) were detected using immunohistochemical staining. In addition, the correlation between the osteoarthritic biomarkers and the Notch2 was also investigated.
 Results: The expression level of Notch signaling were significantly decreased in the postmenopausal osteoarthritic joint, comparing with other groups. No significant differences were observed in all noncollagenase induced joint, regardless of whether being undergone ovariectomy. The expression of Notch2 was negatively correlated to IL-1β and MMP-1 in osteoarthritisoppositely, it was positively correlated to type II collagen.
 Conclusion: the results hinted that estrogen deficiency reduced the expression of Notch signaling in osteoarthritic joint. Thus, the Notch signaling could be a potential target for treating postmenopausal osteoarthritis.

Enhancing autophagy and energy metabolism in the meniscus can delay the occurrence of PTOA in ACLT rat.

Osteoarthritis (OA) is a progressive degenerative joint disease characterized by the destruction of the articular cartilage, meniscus and the like. Autophagy and cellular energy metabolism are the mechanisms by which cells maintain homeostasis. However, little is known about the effects of autophagy and cellular energy metabolism on meniscus degeneration, and the pathogenesis of posttraumatic osteoarthritis (PTOA) after the meniscal injury is rarely reported. Therefore, this study aimed to investigate the relationship between changes in autophagy and cellular energy metabolism in the meniscus following anterior cruciate ligament transection (ACLT) and PTOA induced by subsequent articular cartilage injury. In this study, we use a combination of cell experiments in vitro and animal experiments in vivo. On the one hand, cell experiment results show that inhibiting the mTORC1 signaling pathway by inhibiting the phosphorylation of S6K and AKT proteins in meniscal cells will lead to the increase of Beclin1, LC-3B, ATG12, ULK1, P62, and activate autophagy-related signaling pathways, which in turn protects the extracellular matrix component COL1 of meniscal cells from degradation by catabolic factor MMP13. In addition, it increased the generation of mitochondrial membrane potential in meniscal cells, increased the expression of anti-apoptotic factor BCL-XL, decreased the expression of pro-apoptotic factors BAD and BAX, and reduced the apoptosis of meniscal cells. More importantly, under the stimulation of inflammatory factor IL-1β, the secretion of meniscus cells can reduce the elevated levels of MMP13 and Adamts5 caused by chondrocytes affected by IL-1β. On the other hand, the results of animal experiments in vivo further proved the validity of the results of the cell experiments, and also proved that the meniscus injury did prior to the articular cartilage degeneration after ACLT. In conclusion, this study suggests that the meniscus prior to articular cartilage damage during the development of PTOA after ACLT, and that promoting autophagy and energy metabolism of meniscal cells may be a potential therapeutic target for delaying PTOA.

Comparison of Bone Marrow Mesenchymal Stem Cells-Derived Exosomes, Microvesicles, and Soluble Proteins on Their Chondroprotective Effects and Therapeutic Efficacy of Osteoarthritis Treatment

As a disabling joint disorder, osteoarthritis (OA) is characterized by the degeneration of articular cartilage. Bone marrow mesenchymal stem cells (BMSCs) is considered and used as a tool that is regulated by paracrine for the treatment of OA. Herein, we explored the potential of exosomes (Exos), microvesicles (MVs), and soluble proteins (SPs) produced and isolated from rat BMSCs. First, we observed the biological effects of Exos, MVs, and SPs on IL-1β treated rat chondrocytes. Then, we intra-articularly injected Exos, MVs, SPs, and BMSCs in anterior cruciate ligament transection (ACLT)-induced rat OA model, and observed their therapeutic efficacy. SPs rather than Exos and MVs isolated from rat BMSCs exerted a chondroprotective effect in vitro; MVs and SPs showed comparable effects to the intra-articular injection of BMSCs, whereas the chondroprotective effects of Exos were less potent in vivo. In conclusion, the BMSCs secretome might have inconsistent effects in vivo with in vitro, and Exos might not be superior than other secretome in posttraumatic OA prevention.

Intra-bone marrow injection of magnesium isoglyrrhizinate inhibits inflammation and delays osteoarthritis progression through the NF-κB pathway

ObjectiveOsteoarthritis (OA) presents cartilage damage in addition to chronic inflammation. However, self-recovery of damaged cartilage in an inflammatory environment is not possible. Mesenchymal stem cells (MSCs) in the bone marrow are a source of regenerative repair of damaged cartilage. To date, whether intra-luminal administration of the bone marrow can delay the progression of OA is still unknown. This study, therefore, aimed to explore the role of intra-bone marrow injection of Magnesium isoglycyrrhizinate (MgIG) in delaying the OA progression and to investigate the underlying mechanism.MethodsRabbit OA models were established using the anterior cruciate ligament transection method while a catheter was implanted into the bone marrow cavity. 1 week after surgery, MgIG treatment was started once a week for 4 weeks. The cartilage degradation was analyzed using hematoxylin–eosin staining, Masson’s trichrome staining and Alcian blue staining. Additionally, the pro-inflammatory factors and cartilage regeneration genes involved in the cartilage degeneration and the underlying mechanisms in OA were detected using enzyme-linked immunosorbent assay, quantitative real-time PCR (qRT-PCR) and Western blotting.ResultsThe results of histological staining revealed that intra-bone marrow injection of MgIG reduced degeneration and erosion of articular cartilage, substantially reducing the Osteoarthritis Research Society International scores. Furthermore, the productions of inflammatory cytokines in the bone marrow cavity and articular cavity such as interleukin-1β(IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) were inhibited upon the treatment of MgIG. At the same time, the expression of alkaline phosphate, tartrate-resistant acid phosphatase-5b (TRAP-5b) and C-telopeptides of type II collagen (CTX-II) in the blood also decreased and was positively correlated. On the contrary, cartilage-related genes in the bone marrow cavity such as type II collagen (Col II), Aggrecan (AGN), and SRY-box 9 (SOX9) were up-regulated, while matrix metalloproteinase-3 (MMP-3) was down-regulated. Mechanistically, MgIG was found to exert an anti-inflammatory effect and impart protection to the cartilage by inhibiting the NF-κB pathway.ConclusionIntra-bone marrow injection of MgIG might inhibit the activation of the NF-κB pathway in the progression of OA to exert an anti-inflammatory effect in the bone marrow cavity and articular cavity, thereby promoting cartilage regeneration of MSCs in the bone marrow, making it a potential new therapeutic intervention for the treatment of OA.

P05-14 Effects of a 12-weeks aquatic fitness program in women with osteoarthritis

BackgroundAs a major public health concern, there is a high association between aging and obesity, nutritional deficiencies and physical (in)activity. Thus, diseases related to cartilage are on the list of main concerns of the WHO, assuming the prevention of degeneration of articular cartilage as an important issue for which there are few effective solutions. It is imperative to find preventive strategies that can reduce the incidence of chronic osteoarthritis.MethodsEleven women (58.1±3.3 years-old) diagnosed with knee osteoarthritis (KOA) enrolled in tri-weekly aquatic fitness 45' sessions, for 12 weeks. Taking advantage of the physical properties of the water for increasing the load, a gradual use of the extension of the levers was defined. The warm-up focused on body alignment, joint mobility and breathing. The fundamental part was based on exercises that combine the cardiorespiratory component with the strength component, promoting a superior range of motion. At the end of each session there was a progressive decrease in load, alternating body segments. Before and after the 12 weeks they performed the Senior Fitness Test, hand-grip strength and body measures. All participants were volunteer, informed consent was obtained and all procedures were in accordance to Helsinki Declaration. Sessions were instructed by a CSCS®.ResultsSignificant and meaningful improvements were observed in lower body strength (p > 0.001; d = 1.10), lower body flexibility (p > 0.001; d = 2.88), aerobic endurance (p > 0.001; d = 0.95), dynamic balance (p > 0.001; d = 1.22) and hand grip strength (p > 0.001; d = 1.56). Significant, but moderate improvements were observed in body mass (p = 0.034; d = 0.56) and waist circumference (p = 0.041; d = 0.66).ConclusionsAquatic fitness induced extensive benefits in women conditioning, suggesting that this activity is able to promote an increase in life quality, even if KOA is diagnosed. This program aims to be a tool for implementing healthy behaviour, based on a physical exercise program to people with KOA. With a demographic trend towards an aging population, today society has dragged itself into a vicious cycle to the proven relationship between OA and obesity, and the increasing prevalence of both. To contribute to solving these problems, it is mandatory to have interdisciplinary perspectives that promote a motivating and lasting activity.

Malat1 attenuated the rescuing effects of docosahexaenoic acid on osteoarthritis treatment via repressing its chondroprotective and chondrogenesis activities

Osteoarthritis (OA) is a degenerative disease associated with joint inflammation, articular cartilage degeneration and subchondral hypertrophy. Small molecules which both ameliorate chondrocyte OA phenotype and activate bone marrow-derived mesenchymal stem cells (BMSCs) chondrogenesis under inflammatory conditions have the therapeutical potential for OA treatment. In this study, we characterized a novel small molecule which could ameliorate OA progression via novel regulating mechanisms. Docosahexaenoic acid (DHA), a bioactive molecule, was screened from a small molecule library and showed anti-inflammatory and chondroprotective effects in OA chondrocytes, as well as ameliorated IL-1β impaired BMSCs chondrogenesis in Wnt/β-catenin and NF-κB signaling dependent manners. Furthermore, Malat1 was found to be the key mediator of DHA-mediating anti-inflammation chondroprotection and chondrogenesis. DHA also rescued cartilage loss and damage in a surgery-induced OA mice model. The elevation of serum Malat1 levels caused by OA was also downregulated by DHA treatment. Taken together, our findings demonstrated that DHA, with a dual-signaling repression property, exerted its anti-inflammation, chondroprotection and chondrogenesis function possibly via regulating Malat1 level, suggesting that it may be a possible drug candidate for OA patients with elevated MALAT1 expression levels.

Max interacting protein 1 induces IL-17-producing T helper/regulatory T imbalance in osteoarthritis by upregulating tectonic family member 2

Background/AimOsteoarthritis (OA) is a common total joint disorder associated with regulatory T cell (Treg)/IL-17-producing T helper (Th17) cell imbalance. This study elucidated the mechanism underlying Th17/Treg imbalance during OA progression. MethodsCD4+ T cells were isolated and induced to differentiate and obtain Th17 and Treg cells, and an OA mouse model was established by anterior cruciate ligament transection surgery, followed by loss- and gain-of-function assays. Max interacting protein 1 (MXI1), tectonic family member 2 (TCTN2), Forkhead Box Protein P3 (Foxp3), signal transducer and activator of transcription 3 (STAT3), and retinoic acid receptor-related orphan nuclear receptor gamma t (RORγt) expression was determined in cells and mice, accompanied by the measurement of the proportion of Th17 and Treg cells and the levels of interleukin (IL)− 1β, tumor necrosis factor (TNF)-α, and interferon (INF)-γ. Articular cartilage histopathology was observed by hematoxylin and eosin staining and Safranin O-Fast Green staining. Relationship between MXI1 and TCTN2 was assessed. ResultsBioinformatics analysis identified MXI1 and TCTN2 upregulation in OA patients. Mechanistically, MXI1 bound to TCTN2 promoter to promote its transcription. Upregulated MXI1 boosted INF-γ, STAT3, IL-1β, TNF-α, and RORγt levels and Th17 cell differentiation, but restricted Foxp3 expression and Treg cell differentiation in CD4+ T cells. Effects caused by overexpressed MXI1 were negated by silenced TCTN2. Also, the impacts of MXI1 overexpression on Th17/Treg imbalance and IL-1β, STAT3, TNF-α, Foxp3, INF-γ, and RORγt expression were further validated in OA mice, accompanied by aggravated articular cartilage degeneration. ConclusionConclusively, MXI1 facilitated Th17/Treg imbalance to accelerate OA progression.

Impact of Fluid Flow Shear Stress on Osteoblast Differentiation and Cross-Talk with Articular Chondrocytes.

Bone cells, in particular osteoblasts, are capable of communication with each other during bone growth and homeostasis. More recently it has become clear that they also communicate with other cell-types; including chondrocytes in articular cartilage. One way that this process is facilitated is by interstitial fluid movement within the pericellular and extracellular matrices. This stimulus is also an important mechanical signal in skeletal tissues, and is known to generate shear stresses at the micron-scale (known as fluid flow shear stresses (FFSS)). The primary aim of this study was to develop and characterize an in vitro bone–cartilage crosstalk system, to examine the effect of FFSS on these cell types. Specifically, we evaluated the response of osteoblasts and chondrocytes to FFSS and the effect of FFSS-induced soluble factors from the former, on the latter. This system will ultimately be used to help us understand the role of subchondral bone damage in articular cartilage degeneration. We also carried out a comparison of responses between cell lines and primary murine cells in this work. Our findings demonstrate that primary cells produce a more reliable and reproducible response to FFSS. Furthermore we found that at lower magnitudes , direct FFSS produces anabolic responses in both chondrocytes and osteoblasts, whereas higher levels produce more catabolic responses. Finally we show that exposure to osteoblast-derived factors in conditioned media experiments produced similarly catabolic changes in primary chondrocytes.

Dose-response relationship of in vivo ambulatory load and mechanosensitive cartilage biomarkers-The role of age, tissue health and inflammation: A study protocol.

ObjectiveTo describe a study protocol for investigating the in vivo dose-response relationship between ambulatory load magnitude and mechanosensitive blood markers of articular cartilage, the influence of age, cartilage tissue health and presence of inflammation on this relationship, and its ability to predict changes in articular cartilage quality and morphology within 2 years.DesignProspective experimental multimodal (clinical, biomechanical, biological) data collection under walking stress and three different load conditions varied in a randomized crossover design.Experimental protocolAt baseline, equal numbers of healthy and anterior cruciate ligament injured participants aged 20–30 or 40–60 years will be assessed clinically and complete questionnaires regarding their knee health. Biomechanical parameters (joint kinetics, joint kinematics, and surface electromyography) will be recorded while performing different tasks including overground and treadmill walking, single leg balance and hopping tasks. Magnetic resonance images (MRI) of both of knees will be obtained. On separate stress test days, participants will perform a 30-minute walking stress with either reduced (80% body weight (BW)), normal (100%BW) or increased (120%BW) load. Serum blood samples will be taken immediately before, immediately after, 30, 120 and 210 minutes after the walking stress. Concentration of articular cartilage blood biomarkers will be assessed using enzyme linked immunosorbent assays. At 24-month follow-up, participants will be again assessed clinically, undergo an MRI, complete questionnaires, and have a blood sample taken.ConclusionThe study design provides a standardized set up that allows to better understand the influence of ambulatory load on articular cartilage biomarkers and thereby extend current knowledge on in vivo cartilage metabolism and mechanosensitivity. Further, this study will help to elucidate the prognostic value of the load-induced cartilage biomarker response for early articular cartilage degeneration.Trial registrationThe protocol was approved by the regional ethics committee and has been registered at clinicaltrials.gov (NCT04128566).

Short-term effectiveness of high tibial osteotomy combined with arthroscopic surgery for knee varus arthritis and the results of secondary arthroscopic exploration

To investigate the effectiveness of high tibial osteotomy (HTO) combined with arthroscopic surgery to treat medial compartment knee osteoarthritis (KOA) and secondary arthroscopic exploration to evaluate the outcome of cartilage and meniscus. A clinical data of 57 patients with medial compartment KOA with varus deformity of lower extremities admitted between August 2014 and October 2018 were retrospectively analyzed. There were 23 males and 34 females with an average age of 51.2 years (range, 41-63 years). The disease duration ranged from 2 to 8 years, with an average of 4.7 years. The preoperative femorotibial angle was (179.86±4.69)°, the relative position of the lower limb mechanical axis passing through the tibial plateau was 24.21%±6.98%, and the posterior slope of the tibial plateau was (5.23±1.45)°. The Kellgren-Lawrence grade of knee joint was grade Ⅱ in 22 cases and grade Ⅲ in 35 cases. The preoperative Hospital for Special Surgery (HSS) score, Lysholm score, and visual analogue scale (VAS) score were 59.1±7.3, 48.8±7.6, and 6.2±1.1, respectively. Arthroscopic exploration was performed during the operation to record the articular cartilage degeneration in the weight-bearing area of the medial compartment (Outerbridge grade Ⅰ in 18 cases, grade Ⅱ in 30 cases, and grade Ⅲ in 9 cases) and the condition of the medial meniscus injury, and the corresponding treatment was performed. The coronal force line was adjusted according to the preoperative Kellgren-Lawrence grade of the knee joint during the operation. After operation, the relative position of the lower limb mechanical axis passing through the tibial plateau, the femorotibial angle, and the posterior slope of the tibial plateau were measured; the Kellgren-Lawrence grade of the knee joint was recorded; the Outerbridge grade of articular cartilage degeneration and the meniscus outcome were evaluated by combining with the MRI of the knee joint at 1 year after operation and the second arthroscopic exploration when the internal fixator was removed. The function and pain of the knee were evaluated by Lysholm score, HSS score, and VAS score. All the 57 patients were followed up 36-58 months with an average of 42.1 months. Incisions healed by first intention, and no neurovascular injury, intraarticular or hinge fractures occurred during operation, and no postoperative complications such as deep vein thrombosis of lower limbs and internal fixation failure occurred. All the osteotomy sites healed at 3 months after operation. At 1 year after operation, the internal fixator was removed, and the second arthroscopic exploration showed that there were 15 cases of Outerbridge grade Ⅰ, 31 cases of grade Ⅱ, and 11 cases of grade Ⅲ in the weight-bearing area of the medial compartment, and there was no significant difference when compared with preoperative grade ( Z=31.992, P=0.997); there was no cartilage degeneration in other compartments. Meniscus healing was seen in the injured meniscus, and no injury was seen in the normal meniscus. At last follow-up, there were 19 cases of Kellgren-Lawrence grade Ⅱ and 38 cases of grade Ⅲ, and there was no significant difference when compared with preoperative grade ( Z=49.049, P=0.764). The relative position of the lower limb mechanical axis passing through the tibial plateau was 59.16%±2.87%, and the femorotibial angle was (171.54±3.39)°, which significantly improved when compared with those before operation ( P<0.001). The posterior slope of the tibial plateau was (5.65±1.22)°, which was not significantly different from that before operation ( t=-1.673, P=0.096). The HSS score, Lysholm score, and VAS score were 82.3±7.7, 83.4±6.4, and 1.6±1.1 respectively, which were significantly different from those before operation ( P<0.001). HTO combined with arthroscopic surgery for medial compartment KOA with varus deformity of lower extremities can effectively improve the force line of lower extremities, relieve pain symptoms, and improve joint function, with satisfactory short-term effectiveness, and without significant progress in articular cartilage or meniscus injury after operation.

TGFβ1-modified MSC-derived exosome attenuates osteoarthritis by inhibiting PDGF-BB secretion and H-type vessel activity in the subchondral bone

BackgroundGreater bone resorption increases TGF-β1 release and nestin-positive BMSC recruitment to the subchondral bone marrow, leading to excessive subchondral osteophyte formation and severe wear to articular cartilage. Our previous research demonstrated that BMSCs-ExoTGF-β1 attenuated cartilage damage in osteoarthritis (OA) rats through carrying highly expressed miR-135b. MethodsThe bone marrow mesenchymal stem cells (BMSCs) were isolated from mouse bone marrow, and BMSC-derived exosomes (BMSCs-Exo) were isolated from BMSCs. OA mouse models were established by anterior cruciate ligament transection (ACLT) surgery on the left knee of mice. Then we explored the therapeutic effect of BMSCs-ExoTGF-β1 on ACLT mice. ResultsBMSCs-ExoTGF-β1 attenuated cartilage damage in OA mice in vivo by ameliorating articular cartilage degeneration and suppressing calcification of the cartilage zone. BMSCs-ExoTGF-β1 also inhibited osteoclastogenesis by suppressing the MAPK pathway in vitro. Micro-computed tomography indicated that BMSCs-ExoTGF-β1 impeded uncoupled subchondral bone remodeling. BMSCs-ExoTGF-β1 also reduced CD31hiEmcnhi vessel activity in the subchondral bone and attenuated OA pain behaviors. ConclusionsIn conclusion, BMSCs-ExoTGF-β1 maintains the microarchitecture, inhibits abnormal angiogenesis in subchondral bone and exerts protective effect against OA-induced pain and bone resorption on ACLT mice. Data availabilityThe datasets are available from the corresponding author on reasonable request.

Transplantation of Parathyroid Hormone–Treated Achilles Tendon Promotes Meniscal Regeneration in a Rat Meniscal Defect Model

Background: Autologous tendon grafts are used for meniscal reconstruction of surgically removed knee joint meniscus. However, as meniscal reconstruction cannot prevent the progression of cartilage degeneration, additional procedures that confer meniscus-like histological properties to the transplanted tendon are required for improved outcomes. Hypotheses: Parathyroid hormone (PTH)(1-34) induces cartilage formation in the rat tendon, and transplantation of PTH-treated tendon promotes meniscal regeneration. Study Design: Controlled laboratory study. Methods: Rat Achilles tendon–derived cells were cultured with or without PTH for 28 days and stained with Alcian blue to determine chondrogenic differentiation. After 14 and 28 days of incubation, gene expression was assessed using quantitative real-time polymerase chain reaction. In an in vivo study, rat Achilles tendon was injected with PTH and then transplanted onto a medial meniscal defect. Macroscopic and histological assessments of the regenerated meniscus and of cartilage degeneration in the tibial plateau were performed at 4 and 8 weeks after surgery. Results: In vitro, PTH-treated cells showed better staining with Alcian blue than the control (normal medium) group. PTH1R, Col2a1, Sox9, and RUNX2 were significantly upregulated in PTH-treated cells (P < .05). Macroscopically, the in vivo results revealed more prominent meniscal coverage and lesser progression of articular cartilage degeneration in the PTH group than in the phosphate-buffered saline–injected group. Histologically, toluidine blue staining revealed metachromasia in the PTH-injected tissue at 4 and 8 weeks. The PTH-treated regenerated meniscus showed positive immunostaining for type II collagen in the area exhibiting metachromasia. Moreover, PTH-treated tendon had an enhanced histological score compared with the untreated group at 4 and 8 weeks (P < .05). Conclusion: PTH(1-34) induced cartilage formation in the rat tendon. Transplantation of PTH(1-34)–treated Achilles tendon in a rat meniscal defect model induced meniscal regeneration and preserved knee articular cartilage. Macroscopically, PTH groups showed a greater coverage of the regenerated meniscus. Histologically, the regenerated meniscus had higher cartilaginous matrix content in rats transplanted with PTH-treated tendons. PTH(1-34) stimulated tendon-derived cells to promote chondrogenic differentiation. Clinical Relevance: Meniscal transplantation using PTH-injected autologous tendon grafts might promote meniscal regeneration and prevent progression of cartilage degeneration by stimulating chondrogenic differentiation of tendon-derived cells.

KLF9 and EPYC acting as feature genes for osteoarthritis and their association with immune infiltration

BackgroundOsteoarthritis, a common degenerative disease of articular cartilage, is characterized by degeneration of articular cartilage, changes in subchondral bone structure, and formation of osteophytes, with main clinical manifestations including increasingly serious swelling, pain, stiffness, deformity, and mobility deficits of the knee joints. With the advent of the big data era, the processing of mass data has evolved into a hot topic and gained a solid foundation from the steadily developed and improved machine learning algorithms. Aiming to provide a reference for the diagnosis and treatment of osteoarthritis, this paper using machine learning identifies the key feature genes of osteoarthritis and explores its relationship with immune infiltration, thereby revealing its pathogenesis at the molecular level.MethodsFrom the GEO database, GSE55235 and GSE55457 data were derived as training sets and GSE98918 data as a validation set. Differential gene expressions of the training sets were analyzed, and the LASSO regression model and support vector machine model were established by applying machine learning algorithms. Moreover, their intersection genes were regarded as feature genes, the receiver operator characteristic (ROC) curve was drawn, and the results were verified using the validation set. In addition, the expression spectrum of osteoarthritis was analyzed by immunocyte infiltration and the co-expression correlation between feature genes and immunocytes was construed.ConclusionEPYC and KLF9 can be viewed as feature genes for osteoarthritis. The silencing of EPYC and the overexpression of KLF9 are associated with the occurrence of osteoarthritis and immunocyte infiltration.

Artemisinin relieves osteoarthritis by activating mitochondrial autophagy through reducing TNFSF11 expression and inhibiting PI3K/AKT/mTOR signaling in cartilage

Osteoarthritis (OA) is a widespread chronic degenerative joint disease characterized by the degeneration of articular cartilage or inflamed joints. Our findings indicated that treatment with artemisinin (AT) downregulates the protein levels of MMP3, MMP13, and ADAMTS5, which are cartilage degradation-related proteins in OA, and inhibits the expression of inflammatory factors in interleukin-1β (IL-1β)-stimulated chondrocytes. However, the mechanism of the role of AT in OA remains unclear. Here, we performed gene sequencing and bioinformatics analysis in control, OA, and OA + AT groups to demonstrate that several mRNA candidates were enriched in the PI3K/AKT/mTOR signaling pathway, and TNFSF11 was significantly downregulated after AT treatment. TNFSF11 was downregulated in the OA + AT group, whereas it was upregulated in rat OA tissues and OA chondrocytes. Therefore, we confirmed that TNFSF11 was the target gene of AT. In addition, our study revealed that AT relieved cartilage degradation and defection by activating mitochondrial autophagy via inhibiting the PI3K/AKT/mTOR signaling pathway in IL-1β-induced chondrocytes. Furthermore, an OA model was established in rats with medial meniscus destabilization. Injecting AT into the knee joints of OA rat alleviated surgical resection-induced cartilage destruction. Thus, these findings revealed that AT relieves OA by activating mitochondrial autophagy by reducing TNFSF11 expression and inhibiting PI3K/AKT/mTOR signaling.

Elucidation of the Underlying Mechanism of Gujian Oral Liquid Acting on Osteoarthritis through Network Pharmacology, Molecular Docking, and Experiment.

Gujian oral liquid (GJ), a traditional herbal formula in China, has been widely used to treat patients with osteoarthritis (OA). Nevertheless, the active component and potential mechanism of GJ are not fully elucidated. Thus, we investigate the effect of GJ and explore its underlying mechanism on OA through network pharmacology and experimental validation. First, a total of 175 bioactive compounds were identified, and 134 overlapping targets were acquired after comparing the targets of the GJ with those of OA. 8 hub targets, including IL6 and AKT1, were obtained in PPI network analysis. Then, we built up GJ-target-OA network and protein-protein interaction (PPI) network, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. The results underlined inflammatory tumor necrosis factor (TNF) as a promising signaling pathway of GJ for OA treatment. Moreover, molecular docking also verified the top two active compounds had direct bindings with the top three target genes. Finally, we verified the effect of GJ on OA in vivo and in vitro. In vivo experiments validated that GJ not only significantly attenuated OA phenotypes including articular cartilage degeneration and subchondral bone sclerosis but also reduced the expressions of tumor necrosis factor-α (TNF-α) and p-p65 in articular chondrocytes. Besides, GJ serum also had a protective effect on chondrocytes against inflammation caused by TNF-α in vitro. Hence, our study predicted and verified that GJ could exert anti-inflammation and anticatabolism effects partially via regulating TNF-α/NF-kappa B (NF-κB) signaling.