Cadmium in Market Pork Kidneys: A Study on Cadmium Bioavailability and the Health Effects Based on Mouse Models.

Edible offal of farmed animals can accumulate cadmium (Cd). However, no studies have investigated Cd bioavailability and its health effects. Here, based on mouse models, market pork kidney samples exhibited high Cd relative bioavailability of 74.5 ± 11.2% (n = 26), close to 83.8 ± 7.80% in Cd-rice (n = 5). This was mainly due to high vitamin D3 content in pork kidney, causing 1.7-2.3-fold up-regulated expression of duodenal Ca transporter genes in mice fed pork kidney compared to mice fed Cd-rice, favoring Cd intestinal absorption via Ca transporters. However, although pork kidney was high in Cd bioavailability, subchronic low-dose (5% in diet) consumption of two pork kidney samples having 0.48 and 0.97 µg Cd g-1 dw over 35 d did not lead to significant Cd accumulation in the tissue of mice fed Cd-free rice but instead remarkably decreased Cd accumulation in the tissue of mice fed Cd-rice (0.48 µg Cd g-1) by ∼50% and increased abundance of gut probiotics (Faecalibaculum and Lactobacillus). Overall, this study contributed to our understanding of the bioavailability and health effects associated with Cd in edible offal, providing mechanistic insights into pork kidney consumption safety based on Cd bioavailability.

Baicalein alleviates osteoarthritis by protecting subchondral bone, inhibiting angiogenesis and synovial proliferation.

Osteoarthritis (OA) is one of the most frequent chronic joint diseases with the increasing life expectancy. The main characteristics of the disease are loss of articular cartilage, subchondral bone sclerosis and synovium inflammation. Physical measures, drug therapy and surgery are the mainstay of treatments for OA, whereas drug therapies are mainly limited to analgesics, glucocorticoids, hyaluronic acids and some alternative therapies because of single therapeutic target of OA joints. Baicalein, a traditional Chinese medicine extracted from Scutellaria baicalensis Georgi, has been widely used in anti-inflammatory therapies. Previous studies revealed that baicalein could alleviate cartilage degeneration effectively by acting on articular chondrocytes. However, the mechanisms involved in baicalein-mediated protection of the OA are not completely understood in consideration of integrality of arthrosis. In this study, we found that intra-articular injection of baicalein ameliorated subchondral bone remodelling. Further studies showed that baicalein could decrease the number of differentiated osteoblasts by inhibiting pre-osteoblasts proliferation and promoting pre-osteoblasts apoptosis. In addition, baicalein impaired angiogenesis of endothelial cells and inhibited proliferation of synovial cells. Taken together, these results implicated that baicalein might be an effective medicine for treating OA by regulating multiple targets.

Regulation of TRPV5 transcription and expression by E2/ERα signalling contributes to inhibition of osteoclastogenesis.

The increasing of osteoclasts formation and activity because of oestrogen (E2) deficiency is very important in the aetiology of postmenopausal osteoporosis. Our previous studies showed that E2 inhibited osteoclastic bone resorption by increasing the expression of Transient Receptor Potential Vanilloid 5 (TRPV5) channel. However, the exact mechanism by which E2 increases TRPV5 expression is not fully elucidated. In this study, Western blot, quantitative real-time PCR, tartrate-resistant acid phosphatase staining, F-actin ring staining, chromatin immunoprecipitation and luciferase assay were applied to explore the mechanisms that E2-induced TRPV5 expression contributes to the inhibition of osteoclastogenesis. The results showed that silencing or overexpressing of TRPV5 significantly affected osteoclasts differentiation and activity. Silencing of TRPV5 obviously alleviated E2-inhibited osteoclastogenesis, resulting in increasing of bone resorption. E2 stimulated mature osteoclasts apoptosis by increasing TRPV5 expression. Further studies showed that E2 increased TRPV5 expression through the interaction of the oestrogen receptor α (ERα) with NF-κB, which could directly bind to the fragment of -286 nt ~ -277 nt in the promoter region of trpv5. Taken together, we conclude that TRPV5 plays a dominant effect in E2-mediated osteoclasts formation, bone resorption activity and osteoclasts apoptosis. Furthermore, NF-κB plays an important role in the transcriptional activation of E2-ERα stimulated TRPV5 expression.

Resveratrol Attenuates Lipopolysaccharides (LPS)-Induced Inhibition of Osteoblast Differentiation in MC3T3-E1 Cells.

BACKGROUND LPS-inhibited osteoblastic differentiation plays an important role in the pathogenesis of osteomyelitis. Thus, searching for drugs that affect LPS-mediated osteoblastic differentiation may be crucial in developing therapies for osteomyelitis. The purpose of this study was to investigate the role and mechanisms of resveratrol, a natural polyphenol present in red wine, on LPS-inhibited osteoblastic differentiation. MATERIAL AND METHODS Cell viability was measured by MMT assay. Mitochondrial ATP levels, membrane potential, and superoxide production were measured to evaluate the effects of LPS and resveratrol on mitochondrial functions in osteoblast-like MC3T3-E1 cells. Osteoblast-related genes, including ALP, OCN, OPN, and RUNX2, were measured by ELISA analysis and RT-PCR in differentiated osteoblast cells treated with LPS and resveratrol. Cellular Sirt1 and PCG-1α levels were measured by Western blot to probe the impact of resveratrol treatment in LPS-stimulated MC3T3-E1 osteoblasts. RESULTS The results showed that LPS caused significant mitochondrial dysfunctions of MC3T3-E1 cells in a dose-dependent manner, which were attenuated by resveratrol. Furthermore, LPS markedly decreased the expression of ALP, OCN, OPN, and RUNX2 in MC3T3-E1 cells cultivated in osteoblast differentiation medium, suggesting that LPS inhibited the osteoblastic differentiation of MC3T3-E1 cells. However, resveratrol obviously alleviated the suppressive impact of LPS on osteoblast differentiation. In addition, resveratrol increased expression of Sirt1 and PGC-1α in MC3T3-E1 cells treated with LPS. CONCLUSIONS Taken together, these results show that resveratrol alleviated the suppression of LPS on osteoblast differentiation by improving, at least in part, mitochondrial function.

Regulation of chondrocyte functions by transient receptor potential cation channel V6 in osteoarthritis.

Transient receptor potential vanilloid (TRPV) channels function to maintain the dynamic balance of calcium signaling and calcium metabolism in bones. The goal of this study was to determine the potential role of TRPV6 in regulation of chondrocytes. The level of TRPV6 expression was analyzed by western blot in articular cartilage derived from the knee joints of osteoarthritis (OA) rat models and OA patients. Bone structure and osteoarthritic changes in the knee joints of TRPV6 knockout mice were examined using micro-computed and histological analysis at the age of 6 and 12 months old. Furthermore, to investigate the effects of TRPV6 on chondrocyte extracellular matrix secretion, the release of matrix degrading enzymes, cell proliferation, and apoptosis, we decreased and increased TRPV6 expression in chondrocytes with lentiviral constructs encoding shRNA targeting TRPV6 and encoding TRPV6, respectively. The results showed that the level of TRPV6 expression in an OA rat model was markedly down-regulated. TRPV6 knockout mice showed severe osteoarthritis changes, including cartilage fibrillation, eburnation, and loss of proteoglycans. In addition, deficiency of TRPV6 clearly affected chondrocyte function, such as extracellular matrix secretion, the release of matrix degrading enzymes, cell proliferation, and apoptosis. Taken together, our results implicated that TRPV6 channel, as a chondro-protective factor, was involved in the pathogenesis of OA.

Comparison of the outcome of early and delayed surgical treatment of complete acromioclavicular joint dislocation.

PURPOSE: The aim of this study is to compare the clinical and radiographic results and the complication rate between early and delayed surgical treatment of acromioclavicular joint (ACJ) dislocation. METHODS: Publications in the management of ACJ dislocation are identified from the PubMed database between January 1993 and December 2013 using "acromioclavicular joint" and "dislocation" as keywords. The eligibility criteria included are as follows: (1) ACJ dislocation; (2) intervention, early compared with delayed surgical treatment or the surgical treatment for acute compared with chronic ACJ dislocation; (3) human; and (4) English articles. Exclusion criteria consist of the following: (1) type I and type II ACJ dislocation, (2) no definition of the time of early and delayed surgery in studies, (3) no comparison between the clinical result of early and delayed surgery in studies, (4) laboratory studies, radiographic studies, biomechanical studies, (5) the cases including fractures or revisions in studies, and (6) systematic analyses. RESULTS: Eight studies comparing early and delayed surgical treatment of ACJ dislocation are included in this systematic review. According to Constant scores and shoulder subjective value, early surgery has better functional outcomes than delayed surgery in the treatment of ACJ dislocation (P < 0.05). Partial-dislocation/re-dislocation is found at 26.0 % in early and 38.1 % in delayed surgical treatment (P < 0.05). The rate of CC ossification in early surgical treatment is found as the same as the delayed. The complication rates are found at 12.5 % in early surgical treatment and 17.7 % in the delayed, which is not significantly different. CONCLUSION: Early surgical treatment may have superiority to the delayed procedure in the management of ACJ dislocation with better functional outcomes and more satisfied reduction. However, high-quality evidence studies are required to provide stronger support for this opinion in the future. LEVEL OF EVIDENCE: IV.

Treatment of pelvic fractures through a less invasive ilioinguinal approach combined with a minimally invasive posterior approach.

BACKGROUND: Unstable pelvic fractures usually result from high-energy trauma. There are several treatment modalities available. The purpose of this study was to evaluate the clinical application of a new less invasive ilioinguinal approach combined with a minimally invasive posterior approach technique in patients with unstable pelvic fractures. We also address the feasibility, validity, and limitations of the technique. METHODS: Thirty-seven patients with unstable pelvic fractures were treated with our minimally invasive technique. The anterior pelvic ring fractures were treated with a less invasive ilioinguinal approach, and the sacral fractures were treated with a minimally invasive posterior approach. The clinical outcome was measured using the Majeed scoring system, and the quality of fracture reduction was evaluated. The patients were followed up for 13 to 60 months (mean, 24 months). RESULTS: Anatomical or near to anatomical reduction was achieved in 26 (70.3 %) of the anterior pelvic ring fractures and a satisfactory result was obtained in another 11(29.7 %). For the posterior sacral fractures, excellent reduction was obtained in 33 (89.2 %) of the fractures, with a residual deformity in the other 4 patients. One superficial wound infection and two deep vein thromboses occurred, all of which resolved with conservative treatment. The clinical outcome at one year was "excellent" in 29 patients and "good" in 8 patients (Majeed score). CONCLUSIONS: The satisfactory results showed that a reduction and fixation of unstable pelvic fractures is possible through a combination of a limited ilioinguinal approach and posterior pelvic ring fixation. We believe our method is a new and effective alternative in the management of pelvic fractures.

Estrogen inhibits RANKL-induced osteoclastic differentiation by increasing the expression of TRPV5 channel.

The inhibitor effect of estrogen on osteoclasts differentiation is very important in the etiology of estrogen protecting the adult skeleton against bone loss. However, the precise molecular events underlying the effect of estrogen on osteoclasts differentiation are not known. Recent studies implicated an important role of transient receptor potential vanilloid 5 (TRPV5) in osteoclast differentiation and bone resorption. Furthermore, some studies have confirmed that estrogen is involved in the regulation of calcium ion (Ca(2+)) influx in many cells via TRPV5 channel. Therefore, we hypothesize that TRPV5 channel may be implicated in the process of estrogen-inhibited osteoclastogenesis and bone resorption. Western blot, quantitative real-time PCR, tartrate-resistant acid phosphatase (TRAP) staining, and pit formation assay were employed to investigate the role of TRPV5 in estrogen decreasing osteoclast differentiation and bone resorption. We found that the expression of TRPV5 is significantly down-regulated during estrogen deficiency-induced osteoclastogenesis. Furthermore, TRAP staining and pit formation assay showed that the depletion of TRPV5 significantly blocks the inhibitor effects of estrogen on osteoclasts differentiation and bone resorption activity. Further studies confirmed that estrogen regulates the expression of TRPV5 channel via estrogen receptor. Based on these results above, we can draw conclusion that TRPV5 may contribute to the process of estrogen-inhibited osteoclastogenesis and bone resorption activity.

Knockout of TRPV6 causes osteopenia in mice by increasing osteoclastic differentiation and activity.

BACKGROUND: Calcium ion (Ca(2+)) signals are required for osteoclast differentiation. Previous study showed that transient receptor potential vanilloid 5 (TRPV5) is an essential Ca(2+) transporter in osteoclastogenesis and bone resorption. TRPV5 and TRPV6 represent two highly homologous members within the transient receptor potential (TRP) superfamily. However, the role of TRPV6 in bone metabolism is still controversial and little is known about the involvement of TRPV6 in receptor activator of nuclear factor κ-B ligand (RANKL)-induced osteoclastogenesis. METHODS: In our study, gene knockout mice, RNA interference, western blot, quantitative real-time PCR, tartrate-resistant acid phosphatase (TRAP) staining, pit formation assay, histomorphometry and measurement of serum parameters were employed to investigate the role of TRPV6 in bone homeostasis, osteoclastogenesis and bone resorption. RESULTS: We found that TRPV6 depletion results in noticeable destruction of bone microarchitecture in TRPV6 knockout mice (TRPV6(-/-)), suggesting that TRPV6 is a critical regulator in bone homeostasis. Inactivation of Trpv6 had no effect on osteoblastic bone formation. However, quantification of the TRAP staining showed a significantly increased osteoclast number and surface area in the metaphyseal area of femurs bone sections derived from TRPV6(-/-) mice. In agreement with our observations from TRPV6(-/-) mice, TRPV6 depletion in vitro significantly increased osteoclasts differentiation and bone resorption activity. CONCLUSION: Based on these results above, we can draw conclusions that TRPV6 plays an essential role in bone metabolism and is a critical regulator in osteoclasts differentiation and bone resorption.

Evaluation of the coracoclavicular reconstruction using LARS artificial ligament in acute acromioclavicular joint dislocation.

PURPOSE: The most appropriate procedure for surgical treatment of severe acromioclavicular (AC) joint dislocation was still not clear. The purpose of this study is to evaluate the outcomes of coracoclavicular (CC) reconstruction with ligament augmentation and reconstruction system (LARS) artificial ligaments for the treatment of acute complete AC joint dislocation. METHODS: Twenty-four patients (16 male and 8 female, ages ranged from 21 to 45) with acute complete AC joint dislocations were treated with CC reconstruction using LARS artificial ligaments. All these dislocations were unstable injuries. Clinical evaluation was used by the Constant scores and VAS. The radiographic evaluation consisted of Zanca radiographs for bilateral AC joint and axillary radiographs for the injured shoulder. RESULTS: All patients had follow-up times of 36 months (range 6-60). The Constant scores rose from 62.3 ± 6.9 preoperatively to 94.5 ± 9.3 at final evaluation (P < 0.05). Preoperative VAS scores were 5.1 ± 1.7, and the VAS scores at the last review were 0.7 ± 1.4 (P < 0.05). Follow-up radiographs showed anatomical reduction in 20 patients and slight loss of reduction in 4 patients. Calcification of CC ligament in 4 patients, degenerative change around the AC joint in 2 patient and clavicular osteolysis around screws in one patient were found. CONCLUSIONS: LARS artificial ligament for reconstruction of CC can provide immediate stability and allow early shoulder mobilization with good functional results and few complications. This procedure was an effective and safe method to treat grade III and more AC joint dislocations. LEVEL OF EVIDENCE: IV.

Treatment of distal tibial shaft fractures by three different surgical methods: a randomized, prospective study.

PURPOSE: A few studies focused on the methods of treatment for displaced distal tibial shaft fractures have been published, all of which compared two different methods. In this randomized, prospective study, we aimed to compare minimally invasive plate osteosynthesis, locking intramedullary nail stabilization and external fixation combined with limited open reduction and absorbable internal fixation for distal tibial shaft fractures by assessing complications and secondary procedures. METHODS: From November 2002 to June 2012, 137 skeletally mature patients with displaced distal tibial shaft fractures with or without fibula fracture were randomized to be treated by minimally invasive plate osteosynthesis (group A, n = 46), locking intramedullary nail (group B, n = 46) or external fixation combined with limited open reduction and absorbable internal fixation (group C, n = 45). Age, gender, mechanism of injury, fracture pattern and presence of open fracture were equally distributed among the three groups. Indexes for evaluation included hospital stay, operative time, time to radiographic union, union status, infection and the incidence of re-operation. Mazur ankle score was introduced for functional evaluation. Statistics Analysis System (SAS) 9.2 was used for analysis. RESULTS: A total of 121 patients were included in the final analysis (group A 42, group B 40 and group C 39) and evaluated after a mean of 14.8 months follow-up. There was no significant difference (P > 0.05) in hospital stay, time to radiographic union and the incidence of union status among the three groups. Although group C was associated with less secondary procedures versus groups A and B, it was related with more pin tract infections (15.4 %). Anterior knee pain occurred frequently after locking intramedullary nailing (37.5 %) and the irritation symptoms were more frequently encountered in group A (59.5 %). There was no difference in ankle function between the three methods after operation (P > 0.05). CONCLUSIONS: We consider that the minimally invasive plate osteosynthesis, locking intramedullary nail stabilization and external fixation combined with limited open reduction and absorbable internal fixation techniques are all efficient methods for treating distal tibia fractures. With its wide indications, external fixation combined with limited open reduction and absorbable internal fixation leads to minimal soft tissue complication, good functional result and no local soft tissue irritation or implant removal.

Multistrand titanium cable for the coracoclavicular stabilization of acute acromioclavicular joint dislocation.

The aim of this retrospective study was to evaluate the outcome of the surgical treatment of acute complete acromioclavicular (AC) joint dislocation with multistrand titanium cable for coracoclavicular (CC) stabilization. Forty-two patients with acute complete AC joint dislocation, Rockwood III, IV, V, were treated with CC stabilization using multistrand titanium cable. Thirty-nine patients could be evaluated after a mean follow-up period of 42 months (range, 34-60). The mean VAS score improved from 5.6 +/- 1.5 to 0.4 +/- 1.2 (p < 0.05). The mean Constant score from 64.8 +/- 8.9 preoperatively to 95.3 +/- 9.3 (p < 0.05). Radiographs showed anatomical reduction in 32 out of 39 patients. Cable breakage occurred in 2 patients. CC stabilization with multistrand titanium cables is an effective and safe alternative to other procedures. This procedure provides immediate joint stabilization and allows early mobilization with satisfying functional recovery.

Evaluation of coracoclavicular stabilization of acute acromioclavicular joint dislocation with multistrand titanium cables.

PURPOSE: The aim of this study was to evaluate the outcome of surgical treatment of acute acromioclavicular (AC) joint dislocation with multistrand titanium cables for coracoclavicular (CC) stabilization. METHODS: Forty-two patients with acute AC joint dislocation, including Rockwood type III 14 cases, type IV 2 cases and type V 26 cases, were operated with CC stabilization using multistrand titanium cables. The cables were removed 3-12 months after surgery. The function outcome was evaluated by Constant scores and visual analog scale (VAS) scores. Radiological examination included bilateral antero-posterior and axillary radiography. RESULTS: Three patients were lost to follow-up. Thirty-nine patients had an average follow-up time of 42 months (range 34-60). The Constant scores were 95.3 ± 9.3 at final evaluation. Preoperative and final follow-up VAS scores were 5.6 ± 1.5 and 0.4 ± 1.2, respectively (P < 0.05). Radiographs showed anatomical reduction in 32 patients. Cables breakage occurred in two patients. CONCLUSIONS: CC stabilization with multistrand titanium cables was an effective and safe alternative to other procedures for the treatment of acute high-grade AC joint dislocations. It can provide immediate joint stabilization and allow early mobilization of limb with satisfied functional recovery.

GYY4137-induced p65 sulfhydration protects synovial macrophages against pyroptosis by improving mitochondrial function in osteoarthritis development.

INTRODUCTION: Osteoarthritis (OA) is the most common arthritis that is characterized by the progressive synovial inflammation and loss of articular cartilage. Although GYY4137 is a novel and slow-releasing hydrogen sulfide (H2S) donor with potent anti-inflammatory properties that may modulate the progression of OA, its underlying mechanism remains unclear. OBJECTIVES: In this study, we validated the protective role of GYY4137 against OA pathological courses and elucidated its underlying regulatory mechanisms. METHODS: Cell transfection, immunofluorescence staining, EdU assay, transmission electron microscopy, mitochondrial membrane potential measurement, electrophoretic mobility shift assay, sulfhydration assay, qPCR and western blot assays were performed in the primary mouse chondrocytes or the mouse macrophage cell line raw 264.7 for in vitro study. DMM-induced OA mice model and Macrophage-specific p65 knockout (p65f/f LysM-CreERT2) mice on the C57BL/6 background were used for in vivo study. RESULTS: We found that GYY4137 can alleviate OA progress by suppressing synovium pyroptosis in vivo. Moreover, our in vitro data revealed that GYY4137 attenuates inflammation-induced NLRP3 and caspase-1 activation and results in a decrease of IL-1ß production in macrophages. Mechanistically, GYY4137 increased persulfidation of NF-kB p65 in response to inflammatory stimuli that results in a decrease of cellular reactive oxygen species (ROS) accumulation and ameliorates mitochondrial dysfunctions. Using site-directed mutagenesis, we showed that H2S persulfidates cysteine38 in p65 protein and hampers p65 transcriptional activity, and p65 mutant impaired macrophage responses to GYY4137. CONCLUSION: These findings suggest a mechanism by which GYY4137 through redox modification of p65 participates in inhibiting NLRP3 activation by OA to regulate inflammatory responses. Thus, we propose that GYY4137 represents a promising novel therapeutic strategy for the treatment of OA.

Removal of a broken guide wire entrapped in a fractured femoral neck.

Guide wire plays an important role in the fixation of femoral neck fracture with dynamic hip screw (DHS). Breakage of a guide wire during operation is a very rare condition. We met such a dilemma in DHS fixation of a 54-year-old male patient who sustained Garden type IV fracture of the right femoral neck. The distal end of the guide wire broke and was entrapped in the fractured femoral neck. We tried to get the broken part out by a cannulated drill. Reaming was started with the cannulated drill slowly rotating around the guide K-wire until the reamer fully contained the target under fluoroscope. A bone curette was used to get the broken wire out but failed, so we had to use the cannuated drill to dredge this bone tunnel. Finally the broken wire end was taken out, mixed with blood and bone fragments. Through the existing drilling channel, DHS fixation was easily finished. The patient had an uneventful recovery without avascular necrosis of femoral head or non-union of the fracture at one year's follow-up. A few methods can be adopted to deal with the broken guide wire. The way used in our case is less invasive but technically challenging. When the guide wire is properly positioned, this method is very practical and useful.

Percutaneous lordoplasty for the treatment of severe osteoporotic vertebral compression fractures with kyphosis.

Objective: The study aimed to explore the safety and effectiveness of percutaneous lordoplasty (PLP) in the treatment of severe osteoporotic vertebral compression fracture (OVCF). Methods: Included in this prospective study were patients with single-segment acute severe OVCF who were treated with PLP in our institution from July 2016 to October 2019. Patients' back pain and quality of life were assessed using the visual analog scale (VAS) and SF-36 scores. Lateral X-ray radiography of the spine was performed to measure the vertebral height, vertebral kyphotic angle, and segmental kyphotic angle, and to evaluate the outcome of fracture reduction and kyphotic correction. Intra-and postoperative complications were recorded. Results: Of the 51 included patients, 47 patients were followed up for 12 months. The VAS score decreased from preoperative 7.33 ± 1.92 to postoperative 1.76 ± 0.85 at the 12th month (p < 0.05), and the SF-36 score increased from preoperative 79.50 ± 9.22 to postoperative 136.94 ± 6.39 at the 12th month (p < 0.05). During the 1-year follow-up period, the anterior height of the vertebral body increased significantly from preoperative 10.49 ± 1.93 mm to 19.33 ± 1.86 mm (p < 0.05); the posterior height of the vertebral body increased insignificantly from preoperative 22.23 ± 2.36 mm to 23.05 ± 1.86 mm (p > 0.05); the vertebral kyphotic angle decreased significantly from preoperative 18.33° ± 11.49° to 8.73° ± 1.21° (p < 0.05); and the segmental kyphotic angle decreased significantly from preoperative 24.48° ± 4.64° to 11.70° ± 1.34° (p < 0.05). During the 1-year follow-up period, there was no significant difference in the radiologic parameters, VAS scores, and SF-36 scores, between the 1st day and the 12th month of post-operation (P > 0.05). No nerve damage occurred in any of the cases. Intraoperative cement leakage occurred in six cases, and the fracture of the adjacent vertebral body occurred in one case. Conclusion: PLP can well reduce the risk of fracture and achieve good kyphotic correction and may prove to be a safe, cost-effective and minimally invasive alternative option for the treatment of severe OVCF with kyphotic deformity.

MicroRNA-224-5p nanoparticles balance homeostasis via inhibiting cartilage degeneration and synovial inflammation for synergistic alleviation of osteoarthritis.

MicroRNAs play a crucial role in regulating cartilage extracellular matrix (ECM) metabolism and are being explored as potential therapeutic targets for osteoarthritis (OA). The present study indicated that microRNA-224-5p (miR-224-5p) could balance the homeostasis of OA via regulating cartilage degradation and synovium inflammatory simultaneously. Multifunctional polyamidoamine dendrimer with amino acids used as efficient vector to deliver miR-224-5p. The vector could condense miR-224-5p into transfected nanoparticles, which showed higher cellular uptake and transfection efficiency compared to lipofectamine 3000, and also protected miR-224-5p from RNase degradation. After treatment with the nanoparticles, the chondrocytes showed an increase in autophagy rate and ECM anabolic components, as evidenced by the upregulation of autophagy-related proteins and OA-related anabolic mediators. This led to a corresponding inhibition of cell apoptosis and ECM catabolic proteases, ultimately resulting in the alleviation of ECM degradation. In addition, miR-224-5p also inhibited human umbilical vein endothelial cells angiogenesis and fibroblast-like synoviocytes inflammatory hyperplasia. Integrating the above synergistic effects of miR-224-5p in regulating homeostasis, intra-articular injection of nanoparticles performed outstanding therapeutic effect by reducing articular space width narrowing, osteophyte formation, subchondral bone sclerosis and inhibiting synovial hypertrophy and proliferation in the established mouse OA model. The present study provides a new therapy target and an efficient intra-articular delivery method for improving OA therapy. STATEMENT OF SIGNIFICANCE: Osteoarthritis (OA) is the most prevalent joint disease worldwide. Gene therapy, which involves delivering microRNAs, has the potential to treat OA. In this study, we demonstrated that miR-224-5p can simultaneously regulate cartilage degradation and synovium inflammation, thereby restoring homeostasis in OA gene therapy. Moreover, compared to traditional transfection reagents such as lipofectamine 3000, G5-AHP showed better efficacy in both microRNA transfection and protection against degradation due to its specific surface structure. In summary, G5-AHP/miR-224-5p was developed to meet the clinical needs of OA patients and the high requirement of gene transfection efficiency, providing a promising paradigm for the future application and development of gene therapy.

Urchin-like ceria nanoparticles for enhanced gene therapy of osteoarthritis.

Osteoarthritis (OA) is the most common degenerative joint disease in the world. Gene therapy based on delivering microRNAs (miRNAs) into cells has potential for the treatment of OA. However, the effects of miRNAs are limited by the poor cellular uptake and stability. Here, we first identify a type of microRNA-224-5p (miR-224-5p) from clinical samples of patients with OA that can protect articular cartilage from degeneration and further synthesize urchin-like ceria nanoparticles (NPs) that can load miR-224-5p for enhanced gene therapy of OA. Compared with traditional sphere ceria NPs, the thorns of urchin-like ceria NPs can efficiently promote the transfection of miR-224-5p. In addition, urchin-like ceria NPs have excellent performance of scavenging reactive oxygen species (ROS), which can regulate the microenvironment of OA to further improve the gene treatment of OA. The combination of urchin-like ceria NPs and miR-224-5p not only exhibits favorable curative effect for OA but also provides a promising paradigm for translational medicine.

Isolation, purification and PEG-mediated transient expression of mesophyll protoplasts in Camellia oleifera.

BACKGROUND: Camellia oleifera (C. oleifera) is a woody edible oil crop of great economic importance. Because of the lack of modern biotechnology research, C. oleifera faces huge challenges in both breeding and basic research. The protoplast and transient transformation system plays an important role in biological breeding, plant regeneration and somatic cell fusion. The objective of this present study was to develop a highly efficient protocol for isolating and purifying mesophyll protoplasts and transient transformation of C. oleifera. Several critical factors for mesophyll protoplast isolation from C. oleifera, including starting material (leaf age), pretreatment, enzymatic treatment (type of enzyme, concentration and digestion time), osmotic pressure and purification were optimized. Then the factors affecting the transient transformation rate of mesophyll protoplasts such as PEG molecular weights, PEG4000 concentration, plasmid concentration and incubation time were explored. RESULTS: The in vitro grown seedlings of C. oleifera 'Huashuo' were treated in the dark for 24 h, then the 1st to 2nd true leaves were picked and vacuumed at - 0.07 MPa for 20 min. The maximum yield (3.5 × 107/g·FW) and viability (90.9%) of protoplast were reached when the 1st to 2nd true leaves were digested in the enzymatic solution containing1.5% (w/v) Cellulase R-10, 0.5% (w/v) Macerozyme R-10 and 0.25% (w/v) Snailase and 0.4 M mannitol for 10 h. Moreover, the protoplast isolation method was also applicable to the other two cultivars, the protoplast yield for 'TXP14' and 'DP47' was 1.1 × 107/g·FW and 2.6 × 107/g·FW, the protoplast viability for 'TXP14' and 'DP47' was 90.0% and 88.2%. The purification effect was the best when using W buffer as a cleaning agent by centrifugal precipitation. The maximum transfection efficiency (70.6%) was obtained with the incubation of the protoplasts with 15 µg plasmid and 40% PEG4000 for 20 min. CONCLUSION: In summary, a simple and efficient system for isolation and transient transformation of C. oleifera mesophyll protoplast is proposed, which is of great significance in various aspects of C. oleifera research, including the study of somatic cell fusion, genome editing, protein function, signal transduction, transcriptional regulation and multi-omics analyses.

TRPV6 is a potential regulator of bone resorption in bone loss induced by estrogen deficiency.

The precise effect of estrogen (E2) on osteoclast function is still poorly understood. The aim of this study was to investigate the potential role of transient receptor potential vanilloid 6 (TRPV6) in E2-mediated osteoclast function and to characterize the relevant underlying mechanisms. Here, we found that Trpv6 is drastically decreased in ovariectomy operation animals and the administration of E2 results in an increased expression of Trpv6 in osteoclasts. In contrast, Trpv6 depletion significantly blocked the inhibitory effects of E2 on bone resorption activity, and silencing Trpv6 alleviated E2-induced osteoclast apoptosis. In addition, we found that E2 regulates the transcription of Trpv6 through ERα, by interacting with C/EBPß and NF-κB. Chip assay analysis indicated that C/EBPß regulates Trpv6 transcription by binding to Trpv6 promoter fragments -1,866 nt to -1,761 nt and -2,685 nt to -2,580 nt, whereas NF-κB binds to the -953 nt to -851 nt region. We conclude that TRPV6 has a significant effect on E2-mediated osteoclast function.