LncRNA SNHG1 alleviates myocardial ischaemia-reperfusion injury by regulating the miR-137-3p/KLF4/TRPV1 axis.

AIMS: Myocardial ischaemia-reperfusion injury (MIRI) contributes to serious myocardial injury and even death. Long non-coding RNAs (lncRNAs) have been reported to play pivotal roles in the occurrence and development of MIRI. Here, the detailed molecular mechanism of lncRNA SNHG1 in MIRI was explored. METHODS AND RESULTS: A cell model of MIRI was established through hypoxia/reoxygenation (H/R) stimulation. Cell viability and pyroptosis were evaluated utilizing MTT, PI staining, and flow cytometry. Interleukin (IL)-1ß and IL-18 secretion levels were examined by ELISA. The gene and protein expression were detected by RT-qPCR and western blot, respectively. Dual luciferase reporter gene, RIP and ChIP assays were performed to analyse the molecular interactions. The results showed that lncRNA SNHG1 overexpression alleviated H/R-induced HL-1 cell pyroptosis (all P < 0.05). LncRNA SNHG1 promoted KLF4 expression by sponging miR-137-3p. miR-137-3p silencing alleviated H/R-induced pyroptosis in HL-1 cells (all P < 0.05), which was abolished by KLF4 knockdown (all P < 0.05). KLF4 activated the AKT pathway by transcriptionally activating TRPV1 in HL-1 cells (all P < 0.05). TRPV1 knockdown reversed the alleviation of SNHG1 upregulation on H/R-induced pyroptosis in HL-1 cells (all P < 0.05). CONCLUSIONS: These results showed that lncRNA SNHG1 assuaged cardiomyocyte pyroptosis during MIRI progression by regulating the KLF4/TRPV1/AKT axis through sponging miR-137-3p. Our findings may provide novel therapeutic targets for MIRI.

Surgical technique modification of circumferential decompression for thoracic spinal stenosis and clinical outcome.

Progressive thoracic myelopathy caused by ossification of posterior longitudinal ligament (OPLL) responds poorly to conservative therapy. The most direct decompression is extirpation of ossified posterior longitudinal ligament (PLL). Surgical outcomes of posterior approaches to remove ossified PLL are not always satisfactory because of the risk of neurological deterioration. In this study, we modified the conventional anterior decompression technique via a posterior approach for thoracic OPLL. From an anterior approach, the posterior cortex of vertebral body was exposed and the ossified PLL was removed. Then kyphosis correction was done via posterior instrumentation to reduce cord compression between dura under tension and the anterior canal wall. From the back, the distal end of the ossified PLL was displaced anteriorly to create a gap between ossified PLL and dura, remaining adhesions were divided and the ossified PLL was manipulated through this gap under direct vision. The surgical technique was applied in 20 patients with thoracic myelopathy caused by OPLL. One case of postoperative neurological deterioration was encountered but this recovered fully. Our outcomes were relatively favorable.

Type I collagen scaffold with WNT5A plasmid for in situ cartilage tissue engineering.

BACKGROUND: Cartilage tissue lacks the ability to heal. Cartilage tissue engineering using cell-free scaffolds has been increasingly used in recent years. OBJECTIVE: This study describes the use of a type I collagen scaffold combined with WNT5A plasmid to promote chondrocyte proliferation and differentiation in a rabbit osteochondral defect model. METHODS: Type I collagen was extracted and fabricated into a collagen scaffold. To improve gene transfection efficiency, a cationic chitosan derivative N,N,N-trimethyl chitosan chloride (TMC) vector was used. A solution of TMC/WNT5A complexes was adsorbed to the collagen scaffold to prepare a WNT5A scaffold. Osteochondral defects were created in the femoral condyles of rabbits. The rabbits were divided into defect, scaffold, and scaffold with WNT5A groups. At 6 and 12 weeks after creation of the osteochondral defects, samples were collected from all groups for macroscopic observation and gene expression analysis. RESULTS: Samples from the defect group exhibited incomplete cartilage repair, while those from the scaffold and scaffold with WNT5A groups exhibited "preliminary cartilage" covering the defect. Cartilage regeneration was superior in the scaffold with WNT5A group compared to the scaffold group. Safranin O staining revealed more proteoglycans in the scaffold and scaffold with WNT5A groups compared to the defect group. The expression levels of aggrecan, collagen type II, and SOX9 genes were significantly higher in the scaffold with WNT5A group compared to the other two groups. CONCLUSIONS: Type I collagen scaffold showed effective adsorption and guided the three-dimensional arrangement of stem cells. WNT5A plasmid promoted cartilage repair by stimulating the expression of aggrecan, type II collagen, and SOX9 genes and proteins, as well as inhibiting cartilage hypertrophy.

Wnt5a/Platelet-rich plasma synergistically inhibits IL-1ß-induced inflammatory activity through NF-κB signaling pathway and prevents cartilage damage and promotes meniscus regeneration.

Noncanonical Wnt5a is a particularly attractive growth factor to maintain chondrogenesis. Platelet-rich plasma (PRP) is an autologous blood-derived product and a source of bioactive growth factors involved in tissue regeneration. The present study aimed to investigate the effect and inflammation reaction of Wnt5a/PRP on meniscus cells, and evaluate meniscus regeneration and osteoarthritis (OA) prevention by the application of Wnt5a/PRP gel in a rabbit model of massive meniscal defect. In vitro, the proliferation, migration, differentiation, and interleukin-1 beta (IL-1ß) IL-1ß-induced inflammation reaction of meniscus cells treated by Wnt5a and PRP was assessed. In vivo, the anterior half of the medial meniscus of 18 New Zealand rabbits was excised and implanted with PRP gel, Wnt5a/PRP gel or untreated. After 6 and 12 weeks, the regenerated meniscus were evaluated. Wnt5a can promote the migration of meniscus cells. PRP and Wnt5a had synergistic effect in promoting the proliferation and chondrogenic differentiation of meniscus cells. The IL-1ß-induced meniscus cells study showed that PRP and Wnt5a had the anti-inflammatory actions through nuclear factor kB (NF-κB) signaling pathway. PRP and Wnt5a/PRP significantly inhibited the increase of the p-p65/p65 and p-IκB-α/IκB-α ratios. In vivo transplantation of Wnt5a/PRP gel was demonstrated to promote meniscus regeneration, while reducing OA of knee joint. Wnt5a with PRP had the anti-inflammatory activity in an IL-1ß-induced inflammatory model. They can synergistically improve the chondorgenic differentiation of meniscus cells. Wnt5a/PRP gel treatment could potentially be developed into a new method for meniscus regeneration and the prevention of OA.

The preconditioning of lithium promotes mesenchymal stem cell-based therapy for the degenerated intervertebral disc via upregulating cellular ROS.

Adipose-derived stem cell (ADSC) is one of the most widely used candidate cell for intervertebral disc (IVD) degeneration-related disease. However, the poor survival and low differentiation efficacy in stressed host microenvironment limit the therapeutic effects of ADSC-based therapy. The preconditioning has been found effective to boost the proliferation and the functioning of stem cells in varying pathological condition. Lithium is a common anti-depression drug and has been proved effective to enhance stem cell functioning. In this study, the effects of preconditioning using LiCl on the cellular behavior of ADSC was investigated, and specially in a degenerative IVD-like condition. METHOD: The cellular toxicity on rat ADSC was assessed by detecting lactate dehydrogenase (LDH) production after treatment with a varying concentration of lithium chloride (LiCl). The proliferative capacity of ADSC was determined by detecting Ki67 expression and the relative cell number of ADSC. Then, the preconditioned ADSC was challenged by a degenerative IVD-like condition. And the cell viability as well as the nucleus pulpous (NP) cell differentiation efficacy of preconditioned ADSC was evaluated by detecting the major marker expression and extracellular matrix (ECM) deposit. The therapeutic effects of preconditioned ADSC were evaluated using an IVD degeneration rat model, and the NP morphology and ECM content were assessed. RESULTS: A concentration range of 1-10 mmol/L of LiCl was applied in the following study, since a higher concentration of LiCl causes a major cell death (about 40%). The relative cell number was similar between preconditioned groups and the control group after preconditioning. The Ki67 expression was elevated after preconditioning. Consistently, the preconditioned ADSC showed stronger proliferation capacity. Besides, the preconditioned groups exhibit higher expression of NP markers than the control group after NP cell induction. Moreover, the preconditioning of LiCl reduced the cell death and promoted ECM deposits, when challenged with a degenerative IVD-like culture. Mechanically, the preconditioning of LiCl induced an increased cellular reactive oxidative species (ROS) level and activation of ERK1/2, which was found closely related to the enhanced cell survival and ECM deposits after preconditioning. The treatment with preconditioned ADSC showed better therapeutic effects than control ADSC transplantation, with better NP preservation and ECM deposits. CONCLUSION: These results suggest that the preconditioning with a medium level of LiCl boosts the cell proliferation and differentiation efficacy under a normal or hostile culture condition via the activation of cellular ROS/ERK axis. It is a promising pre-treatment of ADSC to promote the cell functioning and the following regenerative capacity, with superior therapeutic effects than untreated ADSC transplantation.

Restoration of cartilage defects using a superparamagnetic iron oxide-labeled adipose-derived mesenchymal stem cell and TGF-ß3-loaded bilayer PLGA construct.

Aim: We aimed to evaluate the capacity of the bilayer polylactic-co-glycolic acid (PLGA)/TGF-ß3/adipose-derived mesenchymal stem cell (ADSC) construct used to repair cartilage defects and the role of ADSCs in the repair process in vivo. Materials & methods: Defects were created surgically on the femoropatellar groove of knee joints in 64 rabbits. All the rabbits were randomly divided into four groups: defect group, PLGA group, PLGA/TGF-ß3 group and PLGA/TGF-ß3/ADSC group. In vivo MRI and Prussian blue staining were applied. Quantitative real-time PCR and western blot methods were used to analyze the gene and protein expression. Results & conclusion: The result showed that TGF-ß3 could effectively stimulate the expressions of aggrecan, collagen type II and SRY-related HMG box 9 (SOX9). The bilayer PLGA/TGF-ß3/ADSC construct showed a promising repair effect.

An oriented-collagen scaffold including Wnt5a promotes osteochondral regeneration and cartilage interface integration in a rabbit model.

Articular cartilage regeneration remains a major challenge in orthopedics. Noncanonical Wnt5a is a particularly attractive growth factor in this context; Wnt5a inhibits chondrocyte hypertrophy but maintains chondrogenesis. We designed a novel, vertically oriented-collagen scaffold. The effect of Wnt5a on MSCs and chondrocytes and the therapeutic effects of the Wnt5a/oriented-collagen scaffold in terms of osteochondral repair and cartilage integration were evaluated. In vitro, the proliferation, migration, and differentiation of MSCs and chondrocytes treated with Wnt5a, and the mechanisms thereof, were assessed. mRNA microarray analysis was performed to compare the expression profiles of MSCs before and after Wnt5a treatment. In vivo, full-thickness cylindrical osteochondral defects (4 mm in diameter, 3 mm in depth) were created in the patellar grooves of 24 New Zealand white rabbits and implanted with oriented-collagen scaffolds (n = 8), Wnt5a/oriented-collagen scaffolds (n = 8), or nothing (n = 8). After 6 and 12 weeks, integration and tissue responses were evaluated. The proliferation, migration, chondrogenic differentiation, and extracellular matrix formation of/by MSCs and chondrocytes improved greatly after treatment with Wnt5a. Western blotting showed that the PI3K/AKT/JNK signaling pathway was activated. Microarray analysis revealed that the Wnt5a group exhibited a significant upregulation of the PI3K pathway. Reactome GSEA pathway interaction analysis revealed that such upregulation was associated with collagen and extracellular matrix formation. In vivo, the Wnt5a/oriented-collagen scaffold group exhibited optimal interface integration, cartilage regeneration, and collagenous fiber arrangement, accompanied by significantly increased glycosaminoglycan and collagen accumulations in the zones of regeneration and integration, compared to the other groups. Gene expression analysis showed that the levels of mRNAs encoding genes involved in cartilage formation were significantly increased in the Wnt5a/oriented, collagen scaffold group (all P < .05). Wnt5a promoted the proliferation, migration, and chondrogenic differentiation of MSCs and chondrocytes via the activation of the PI3K/AKT/JNK signaling pathway. The Wnt5a/oriented-collagen constructs enhanced the structure-specific regeneration of hyaline cartilage in a rabbit model and may be a promising treatment for the repair of human cartilage defects.

A case report of atypical long segmental thoracic hypertrophic pachymeningitis with ossification of ligamentum flavum and literature review.

BACKGROUND: Thoracic spinal stenosis caused by ossification of ligamentum flavum (OLF) is frequently seen, but long segmental thoracic spinal cord compressed by consequent thickened ligament posteriorly was rarely reported. OBJECT: To report a case of thoracic spinal cord compression caused by atypical long segmental thoracic hypertrophic pachymeningitis (HP) with OLF. METHODS: A 55-year-old woman presenting with weakness and numbness in lower extremities was admitted to our department. Combined with physical examination and MRI results, diagnosis of HP with OLF was considered. Due to progressive neurological symptoms, thoracic decompression with internal fixation was performed. RESULTS: The patient felt a reduced numbness and improvement in motor functions 5 days after surgery. Pathological examination suggested the diagnosis of HP with OLF. CONCLUSIONS: HP is a rare condition characterized as thickening and enhancement of the dura mater on contrast-enhanced MRI and chronic inflammatory hyperplasia changes on biopsy. A case of atypical HP complicated with OLF is described. Chondrocytes infiltration in histological examination indicates the potential of ossification in HP.