Leptin differentially regulate cell apoptosis and cycle by histone acetylation in tibial and vertebral epiphyseal plates.

Leptin showed different apoptosis regulation effects on the chondrocytes from tibial and vertebral epiphyseal plates. However, the mechanism is still unclear. In this study, we tested the protein profile of tibial and vertebral epiphyseal plate chondrocytes with and without leptin stimulation by mass spectrometry and found that the histone acetylation level of tibial chondrocytes was decreased after leptin treatment, while increased in vertebral epiphyseal plates. COIP assay showed that leptin promoted H3, H4 histone acetylation by recruiting CREB binding protein (CBP)/P300 to activate histone acetyl transferases (HATs) activity in vertebral disc chondrocytes. But in tibial plate cartilage cells, leptin did not recruit CBP and p300, thus differently affect the apoptosis of epiphyseal plate chondrocytes. Through explored the mechanism of histone acetylation modulated by leptin, and its effect on cartilage cell apoptosis and cell cycle regulation, This provides a novel target therapy possibility therapeutic approach to for the related disease.

Posterior cruciate ligament tibial attachment sacrifice percentage is higher in cruciate-retaining total knee arthroplasty in patients with discoid lateral meniscus.

BACKGROUND: The posterior cruciate ligament (PCL) attachment may be damaged in cruciate-retaining total knee arthroplasty (CR-TKA) using the complete resection for tibial preparation, and resection amount varies greatly among individuals. Discoid lateral meniscus (DLM) is one of the most common anatomic knee variants. This study aimed to evaluate the difference in PCL attachment sacrifice in CR-TKA between patients with and without DLM. METHODS: Fifty-one knees in the study group (DLM group) were matched 1:1 to 51 control knees (non-DLM group) by age, sex, and maximum width of the tibial plateau. The percentage of the sacrificed PCL attachment and the morphological parameters of the tibial plateau were evaluated using magnetic resonance imaging (MRI) in a blind manner. RESULTS: With a tibial cut simulated at a 0°, 3°, and 7° osteotomy slope, the mean PCL attachment resection percentages in the non-DLM group were 40.5%, 53.6%, and 72.6%, respectively. The corresponding resection percentages in the DLM group were 61.0% (P < 0.001), 73.3% (P < 0.001), and 85.7% (P < 0.001), respectively. The percentage of the minimum meniscus width to the maximum tibia width showed a weak positive correlation with the percentage of PCL attachment sacrifice. CONCLUSIONS: A significantly greater portion of PCL attachment was sacrificed in DLM patients undergoing CR-TKA using the complete proximal tibia resection. Attention should be paid to PCL attachment resection during CR-TKA in patients with DLM, and alternative techniques or prosthesis types should be considered.

Leptin accelerates BMSC transformation into vertebral epiphyseal plate chondrocytes by activating SENP1-mediated deSUMOylation of SIRT3.

Bone marrow mesenchymal stem cells (BMSCs) are capable of multidirectional differentiation, and engrafted BMSCs can be used to replace damaged chondrocytes for treatment of intervertebral disc disease. However, chondroblast differentiation of implanted BMSCs is inhibited by the anoxic environment of the articular cavity. Here, we found that leptin enhanced the transformation of BMSCs into chondrocytes under hypoxic conditions. BMSCs isolated from mice were cultured in medium supplemented with leptin under hypoxia. The expression of MFN1/2 and OPA1 were increased only in BMSCs cultured in an anoxic environment. In addition, in hypoxic environments cell energy metabolism relies on glycolysis regulated by leptin, rather than by mitochondrial oxidation. The expression of the de-SUMOylation protease SENP1 was elevated, leading to SIRT3-mediated activation of PGC-1α; these processes were regulated by CREB phosphorylation, and promoted mitochondrial fusion and cell differentiation. The chondrogenic activity of BMSCs isolated from SIRT3-knockout mice was lower than that of BMSCs isolated from wildtype mice. Implantation of SIRT3-knockout murine-derived BMSCs did not significantly improve the articular cartilage layer of the disc. In conclusion, the hypoxic microenvironment promoted BMSC differentiation into chondrocytes, whereas osteoblast differentiation was inhibited. SENP1 activated SIRT3 through the deSUMOylation of mitochondria and eliminated the antagonistic effect of SIRT3 acetylation on phosphorylation. When phosphorylation activity of CREB was increased, phosphorylated CREB is then transferred to the nucleus, affecting PGC-1α. This promotes mitochondrial fusion and differentiation of BMSCs. Leptin not only maintains chondrogenic differentiation homeostasis of BMSCs, but also provides energy for differentiation of BMSCs under hypoxic conditions through glycolysis.

Recent Advances in Multifunctional Hydrogels for the Treatment of Osteomyelitis.

Osteomyelitis (OM), a devastating disease caused by microbial infection of bones, remains a major challenge for orthopedic surgeons. Conventional approaches for prevention and treatment of OM are unsatisfactory. Various alternative strategies have been proposed, among which, hydrogel-based strategies have demonstrated potential due to their unique properties, including loadable, implantable, injectable, printable, degradable, and responsive to stimuli. Several protocols, including different hydrogel designs, selection of antimicrobial agent, co-administration of bone morphogenetic protein 2 (BMP 2), and nanoparticles, have been shown to improve the biological properties, including antimicrobial effects, osteo-induction, and controlled drug delivery. In this review, we describe the current and future directions for designing hydrogels and their applications to improve the biological response to OM in vivo.

The local concentration of Ca2+ correlates with BMP7 expression and osseointegration in patients with total hip arthroplasty.

BACKGROUND: A successful osseointegration of total hip arthroplasty (THA) relies on the interplay of implant surface and bone marrow microenvironment. This study was undertaken to investigate the impact of perioperative biochemical molecules (Ca2+, Mg2+, Zn2+, VD, PTH) on the bone marrow osteogenetic factors (BMP2, BMP7, Stro-1+ cells) in the metaphyseal region of the femoral head, and further on the bone mineral density (BMD) of Gruen R3. METHODS: Bone marrow aspirates were obtained from the discarded metaphysis region of the femoral head in 51 patients with THA. Flow cytometry was used to measure the Stro-1+ expressing cells. ELISA was used to measure the concentrations of bone morphologic proteins (BMP2 and BMP7) and the content of TRACP5b in serum. TRAP staining was used to detect the osteoclast activity in the hip joint. The perioperative concentrations of the biochemical molecules above were measured by radioimmunoassay. The BMD of Gruen zone R3 was examined at 6 months after THA, using dual-energy X-ray absorptiometry (DEXA). RESULTS: Our data demonstrated that the concentration of Ca2+ was positively correlated with BMP7 expression, and with the postoperative BMD of Gruen zone R3. However, the concentration of Mg2+ had little impact on the R3 BMD, although it was negatively correlated with the expression of BMP7. Osteoclast activity in hip joint tissue of patients with femoral neck fractures was increased. Compared with the patients before THA, the levels of TRACP5b in serum of patients after THA were decreased. The data also suggested that the other biochemical molecules, such as Zn2+, VD, and PTH, were not significantly correlated with any bone marrow osteogenetic factors (BMP2, BMP7, Stro-1+ cells). The postoperative R3 BMD of patients of different gender and age had no significant difference. CONCLUSIONS: These results indicate the local concentration of Ca2+ may be an indicator for the prognosis of THA patients.

The effect of trace elements on BMP-2, BMP-7 and STRO-1+ cells in hip replacement.

To explore the correlation between the trace elements in the proximal femur and BMP-2, BMP-7 and STRO-1+ cells in hip replacement, and analyze the therapeutic effect of prosthesis loosening in clinic. Fifty-one patients undergone the first hip replacement in xxx hospital from August 2016 to August 2019 were selected as the study subjects, including 26 females and 25 males, aged 52-89 years. The bone marrow mesenchymal stem cells (BMSCs) were cultured in vitro for flow cytometry, and the string-1+ in BMSCs was detected and analyzed. After that, the expression of bone morphogenetic protein 2 (BMP-2) and bone morphogenetic protein 7 (BMP-7) in the cells were detected by enzyme-linked immunosorbent assay, the content of trace elements in the supernatant was detected by radioimmunoassay, and the collected data were analyzed statistically. In the analysis of the content of trace elements, it was found that the correlation between trace elements was dependent on the separation area, and all trace elements had no correlation with BMP2. Ca2+, Mg2+ were correlated with the level of BMP7 and Ca2+, VD3 was correlated with the percentage of STOR-1+ cells. Further analysis showed that the correlation between trace elements was dependent on bone mineral density (BMD) area, and there was a positive correlation between vitamin D3 (VD3), parathyroid hormone (PTH), zinc, and BMD in zone 7. To sum up, it is found that trace elements may be related to prosthesis loosening, which provides experimental basis for the treatment of prosthesis loosening later.

Overexpression of miR­146a blocks the effect of LPS on RANKL­induced osteoclast differentiation.

The concept that inflammation serves a leading role in osteoclast­induced bone loss under pathological circumstances is now widely accepted. In the present study, it was observed that lipopolysaccharides (LPSs) demonstrated a synergic effect on receptor activator of nuclear factor κ­B ligand (RANKL)­induced osteoclast differentiation in Raw264.7 cells, with increasing levels of multiple pro­inflammatory cytokines including interleukin (IL)­6, tumor necrosis factor­α and IL­1ß. Furthermore, microRNA (miR)­146a was highly induced by LPS and RANKL co­stimulation during the process of osteoclast differentiation. Overexpression of miR­146a inhibited osteoclast transformation by targeting the key regulators of nuclear factor (NF)­κß signaling, TNF receptor­associated factor 6 and interleukin­1 receptor­associated kinase 1. The downstream activation of NF­κß signaling was also inhibited by transfection with a miR­146a mimic. Altogether, the results of the present study demonstrated that miR­146a prevents osteoclast differentiation induced by LPS and RANKL co­stimulation, suggesting that miR­146a may be a promising therapeutic target for treatment of inflammation mediated bone loss.

Expression of tissue inhibitor of metalloproteinases-1 and B-cell lymphoma-2 in the synovial membrane in patients with knee osteoarthritis.

The purpose of this study was to determine the expression and impact of tissue inhibitor of metalloproteinases-1 (TIMP-1) and B-cell lymphoma-2 (Bcl-2) in knee osteoarthritis (KOA). We collected synovial fluids from the knee joint of 70 KOA patients and 30 controls. The expression levels of TIMP-1 and Bcl-2 were significantly higher in KOA patients than those in the control group (P<0.01). We also found positive correlation between the severity of KOA and the expression level of TIMP-1 (r=0.8027, P<0.05) and and Bcl-2 (r=0.5336, P<0.05). However, we found no correlation between the expression levels of TIMP-1 and Bcl-2 in the synovial membranes of KOA patients (P>0.05). Both TIMP-1 and Bcl-2 are expressed at high levels in the synovial membrane with KOA, and are closely related to the occurrence and development of KOA. Thus, detection of TIMP-1 and Bcl-2 in KOA patients can be helpful in diagnosing the state of KOA.

Small interfering RNA-mediated silencing of G-protein-coupled receptor 137 inhibits growth of osteosarcoma cells.

PURPOSE: Osteosarcoma is the most widespread primary carcinoma in bones. Osteosarcoma cells are highly metastatic and frequently develop resistance to chemotherapy making this disease harder to treat. This identifies an urgent need of novel therapeutic strategies for osteosarcoma. G-Protein-coupled receptor 137 (GPR137) is involved in several human cancers and may be a novel therapeutic target. METHODS: The expression of GPR137 was assessed in one osteoblast and three human osteosarcoma cell lines via the quantitative real-time polymerase chain reaction and western blot assays. Stable GPR137 knockdown cell lines were established using an RNA interference lentivirus system. Viability, colony formation, and flow cytometry assays were performed to measure the effects of GPR137 depletion on cell growth. The underlying molecular mechanism was determined using signaling array analysis and western blot assays. RESULTS: GPR137 expression was higher in the three human osteosarcoma cell lines, Saos-2, U2OS, and SW1353, than in osteoblast hFOB 1.19 cells. Lentivirus-mediated small interfering RNA targeting GPR137 successfully knocked down GPR137 mRNA and protein expression in both Saos-2 and U2OS cells. In the absence of GPR137, cell viability and colony formation ability were seriously impaired. The extent of apoptosis was also increased in both cell lines. Moreover, AMP-activated protein kinase α, proline-rich AKT substrate of 40 kDa, AKT, and extracellular signal-regulated kinase phosphorylation levels were down-regulated in GPR137 knockdown cells. CONCLUSIONS: The results of this study highlight the crucial role of GPR137 in promoting osteosarcoma cell growth in vitro. GPR137 could serve as a potential therapeutic target against osteosarcoma.