Piezo1 mediates the degradation of cartilage extracellular matrix in malocclusion-induced TMJOA.

OBJECTIVES: To evaluate the role of Piezo1 in the malocclusion-induced osteoarthritic cartilage of the temporomandibular joint. METHODS: A temporomandibular joint osteoarthritis model was established using a unilateral anterior crossbite in vivo, and cartilage degeneration and Piezo1 expression were observed by histological and immunohistochemical staining. ATDC5 cells were loaded with 24 dyn/cm2 fluid flow shear stress using the Flexcell device in vitro and expression and function of Piezo1 were evaluated. After identifying the function of Piezo1 in YAP translocation under FFSS conditions, the influence of Piezo1 and YAP on metabolism-related enzymes under FFSS was detected through a real-time polymerase chain reaction analysis and western blotting. A UAC-TMJ injection model was established to observe the therapeutic effect of intra-articular injection of a Piezo1 inhibitor on osteoarthritic cartilage matrix loss. RESULTS: Piezo1 was overexpressed in the osteoarthritic cartilage and cultured chondrocytes under shear stress. Piezo1 Silencing inhibited the nuclear translocation of YAP and subsequently downregulated the expression of MMP13 and ADAMTS5. Intra-articular injection of the Piezo1 inhibitor, GsMTx4, could ameliorate proteoglycan degradation in malocclusion-induced TMJOA and suppressed MMP13 and ADAMTS5 expression. CONCLUSIONS: Our results revealed that the activation of Piezo1 promotes mechanical-induced cartilage degradation through the YAP-MMP13/ADAMTS5 signaling pathway.

[Oral Squamous Cell Carcinoma-Derived Cell-Free DNA Modulates Stemness and Migration of Oral Squamous Cell Carcinoma Cell Line by Inducing M2 Macrophage Polarization].

Objective: To investigate the effect of oral squamous cell carcinoma (OSCC)-derived cell-free DNA (cfDNA) on the polarization of macrophages and the regulatory effect of polarized macrophages on the stemness and migration of OSCC cells. Methods: A total of 30 OSCC tissue samples, 10 dysplastic oral tissue samples, and 10 normal oral tissue samples were collected. The status of all tissue samples was confirmed by pathology analysis. Immunohistochemical (IHC) staining and immunofluorescence (IF) staining were performed to examine the cell count and location of M2 macrophages in different types of oral tissue samples. The conditioned medium (CM) of OSCC cell line CAL-27 from the human tongue was collected and the cfDNA was concentrated and isolated for identification. The macrophages were treated by cfDNA and their morphological characteristics were observed under microscope. The expression levels of polarization-related indicators were determined by RT-qPCR. CAL-27 cell line was treated with macrophage CM induced by cfDNA and the expression levels of stemness-related genes were determined by RT-qPCR. Scratch-wound assay was conducted to verify that the migration ability of CAL-27 was modulated by macrophages induced by cfDNA. Results: There were more M2 macrophages in the deep connective tissue of dysplastic oral epithelium and the stroma of OSCC compared with those in the normal oral tissues ( P<0.05). OSCC cell line CAL-27 could secret cfDNA of 10000-15000 bp in length. cfDNA secreted by CAL-27 could induced in macrophages significantly higher expression of M2-macrophage-related genes ( P<0.05). cfDNA-treated macrophages induced significantly increased expression of stemness-related genes in CAL-27 cell line ( P<0.05) and promoted the migration ability of CAL-27 cell line ( P<0.05). Conclusion: OSCC-derived cfDNA promotes stemness and migration of OSCC cell line by inducing M2 macrophage polarization.

Applicative Assessment of a Selective Laser Melting 3D-Printed Ti-6Al-4 V Plate with a Honeycomb Structure in the Reconstruction of a Mandibular Defect of a Beagle Dog.

The most challenging problem in oral and maxillofacial surgery is the reconstruction of defects for the oral and maxillofacial complex. Transfer of different autografts is known as the "gold standard" for the reconstruction of bone defects in the oral and maxillofacial region. Graft harvesting, however, can lead to many complications, such as donor-site morbidity, surgical time-consuming, etc. Three-dimensional (3D) printing technology is an innovative technique that allows the fabrication of personalized plates and scaffolds to fit the precise anatomy of an individual's defect. In this study, a selective laser melting 3D-printed Ti-6Al-4 V plate with a honeycomb was designed, and its physical and biological features were characterized. The personalized 3D-printed scaffold and commercialized titanium reconstruction plate were applied to reconstruct a 4 cm mandibular defect in a beagle dog. Effects of the treatment were analyzed radiologically and histologically. Our results showed that the application of a 3D-printed plate with a honeycomb achieved good biocompatibility and osseointegration and has potential clinical application.

Effect of air-blowing temperature and water storage time on the bond strength of five universal adhesive systems to dentin.

The purpose of this study was to evaluate the air-blowing temperature and water storage time on the micro-tensile bond strength (µTBS) of five universal adhesive systems to dentin. The bond strength with two different air-blowing temperatures (60±2ºC and 23±2ºC) was measured after water storage at 37ºC for 24 h and 100 days respectively. The fracture surface on dentin side was observed by scanning electron microscope (SEM). Three-way ANOVA revealed a significant effect of universal system (p<0.001) and air-blowing temperature (p<0.001) on bond strength to dentin except water-storage time (p=0.145). The interaction within three factors was significantly different (p<0.001). It could be concluded that the µTBS of universal systems to dentin was material-depended. The higher and more stable bonding performance of universal systems on dentin could be achieved by air-blowing at 60±2ºC temperature. In addition, the quantity of voids in the adhesive layer of aceton-based universal adhesive was significantly reduced by higher temperature.

Metformin Carbon Dots for Promoting Periodontal Bone Regeneration via Activation of ERK/AMPK Pathway.

The osteogenic potential of mesenchymal stem cells (MSCs) is severely impaired under persistent inflammation of periodontitis. A highly efficient way to promote or rescue osteogenic potential of MSCs under inflammation remains an unmet goal. Herein, metformin carbon dots (MCDs) with excellent biocompatibility are prepared from metformin hydrochloride and citric acid via a hydrothermal method. The MCDs can more effectively enhance the alkaline phosphatase (ALP) activity, calcium deposition nodules formation, expression of osteogenic genes and proteins in rat bone marrow mesenchymal stem cells (rBMSCs) than metformin under both inflammatory and normal conditions. Moreover, a novel pathway of extracellular signal-regulated kinases (ERK)/AMP-activated protein kinase (AMPK) signaling is involved in the MCDs-induced osteogenesis. In periodontitis rats, MCDs can effectively regenerate the lost alveolar bone, but not the metformin. Taken together, MCDs can be the promising candidate nanomaterial for periodontitis treatment. This work may provide a new pharmacological target of ERK/AMPK pathway for treating bone loss and also give additional insights into developing nanodrugs from the numerous medications.

Bone mesenchymal stem cells are recruited via CXCL8-CXCR2 and promote EMT through TGF-ß signal pathways in oral squamous carcinoma.

OBJECTIVES: Bone mesenchymal stem cells (BMSCs) play critical roles in tumour microenvironment. However, molecular mechanisms of how BMSCs to be recruited and effect subsequent tumour progression are poorly understood in oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: The distribution of CXCL8 was detected by immunohistochemical staining in OSCC tissues. The chemotaxis of conditioned media from different epithelial cells to BMSCs was examined by trans-well assay. Real-time quantitative PCR (qPCR) and ELISA were used to detect the expression of related cytokines and chemokine receptors. The migration of BMSCs was observed in BALB/c nude mice. The roles of BMSCs in proliferation, migration and invasion of OSCC were detected by CCK-8, flow cytometry and trans-well assay. Epithelial-mesenchymal transition (EMT)-related markers were analysed by qPCR and Western blot in vitro, and growth was evaluated in BALB/c nude mice using subcutaneously implanted OSCC in nude mouse model in vivo. RESULTS: Using OSCC, we show CXCL8, secreted by OSCC, binds to exclusively CXCR2 in BMSCs to facilitate migration of BMSCs to OSCC. TGF-ß secreted by BMSCs subsequently induces EMT of OSCC to promote their proliferation, migration and infiltration. We also showed that the Ras/Raf/Erk axis plays a critical role in tumour progression. CONCLUSIONS: Our results provide the molecular basis for BMSC recruitment into tumours, and how this process leads to tumour progression and leads us to develop a novel OSCC treatment target.