Mitochondria-specific antioxidant MitoTEMPO alleviates senescence of bone marrow mesenchymal stem cells in ovariectomized rats.

Senescence in bone marrow mesenchymal stem cells (BMSCs), triggered by excessive oxidative stress, plays a crucial role in the onset of postmenopausal osteoporosis. Recent studies underscore the importance of mitochondrial rehabilitation and quality control as key determinants in the modulation of oxidative stress and cellular senescence. MitoTEMPO, a mitochondria-targeted antioxidant, has been shown to mitigate the heightened levels of reactive oxygen species (ROS). In our research, we observed that BMSCs from ovariectomized (OVX) rats displayed premature senescence, which was attributed to combined mitochondrial and lysosomal dysfunction, a condition that worsens with extended estrogen deprivation. Treatment with MitoTEMPO effectively reversed these effects, reinstating lysosomal functionality and suppressing the mitochondrial unfolded protein response (UPRmt). Subsequent in vivo experiments corroborated these observations, revealing that MitoTEMPO administration in OVX rats curtailed trabecular bone loss and reduced the expression of p53, HSP60, and CLPP in the trabecular bone region of the proximal tibia. Overall, our findings suggest that MitoTEMPO holds promise as a therapeutic agent to counteract senescence in OVX-BMSCs, offering a potential strategy for treating postmenopausal osteoporosis.

Uncovering the shared neuro-immune-related regulatory mechanisms between spinal cord injury and osteoarthritis.

Adults with spinal cord injury (SCI), a destructive neurological injury, have a significantly higher incidence of osteoarthritis (OA), a highly prevalent chronic joint disorder. This study aimed to dissect the neuroimmune-related regulatory mechanisms of SCI and OA using bioinformatics analysis. Using microarray data from the Gene Expression Omnibus database, differentially expressed genes (DEGs) were screened between SCI and sham samples and between OA and control samples. Common DEGs were used to construct a protein-protein interaction (PPI) network. Weighted gene co-expression network analysis (WGCNA) was used to mine SCI- and OA-related modules. Shared miRNAs were identified, and target genes were predicted using the Human MicroRNA Disease Database (HMDD) database. A miRNA-gene-pathway regulatory network was constructed with overlapping genes, miRNAs, and significantly enriched pathways. Finally, the expression of the identified genes and miRNAs was verified using RT-qPCR. In both the SCI and OA groups, 185 common DEGs were identified, and three hub clusters were obtained from the PPI network. WGCNA revealed three SCI-related modules and two OA-related modules. There were 43 overlapping genes between the PPI network clusters and the WGCNA network modules. Seventeen miRNAs shared between patients with SCI and OA were identified. A regulatory network consisting of five genes, six miRNAs, and six signaling pathways was constructed. Upregulation of CD44, TGFBR1, CCR5, and IGF1, while lower levels of miR-125b-5p, miR-130a-3p, miR-16-5p, miR-204-5p, and miR-204-3p in both SCI and OA were successfully verified using RT-qPCR. Our study suggests that a miRNA-gene-pathway network is implicated in the neuroimmune-related regulatory mechanisms of SCI and OA. CD44, TGFBR1, CCR5, and IGF1, and their related miRNAs (miR-125b-5p, miR-130a-3p, miR-16-5p, miR-204-5p, and miR-204-3p) may serve as promising biomarkers and candidate therapeutic targets for SCI and OA.

Metabolic profiling of synovial fluid in human temporomandibular joint osteoarthritis.

Introduction: Temporomandibular joint (TMJ) osteoarthritis (OA) is a common TMJ degenerative disease with an unclear mechanism. Synovial fluid (SF), an important component of TMJ, contains various proteins and metabolites that may directly contribute to OA. The present study aimed to investigate the influence of SF in TMJOA at the metabolite level. Methods: Untargeted and widely targeted metabolic profiling were employed to identify metabolic changes in SF of 90 patients with different TMJOA grades according to TMJ magnetic resonance imaging. Results: A total 1498 metabolites were detected. Most of the metabolites were amino acids and associated metabolites, benzene and substituted derivatives, and lipids. Among patients with mild, moderate and severe TMJOA, 164 gradually increasing and 176 gradually decreasing metabolites were identified, indicating that biosynthesis of cofactors, choline metabolism, mineral absorption and selenocompound metabolism are closely related to TMJOA grade. Combined metabolomics and clinical examination revealed 37 upregulated metabolites and 16 downregulated metabolites in patients with pain, of which 19 and 26 metabolites were positively and negatively correlated, respectively, with maximum interincisal opening. A model was constructed to diagnose TMJOA grade and nine biomarkers were identified. The identified metabolites are key to exploring the mechanism of TMJOA. Discussion: In the present study, a metabolic profile was constructed and assessed using a much larger number of human SF samples from patients with TMJOA, and a model was established to contribute to the diagnosis of TMJOA grade. The findings expand our knowledge of metabolites in human SF of TMJOA patients, and provide an important basis for further research on the pathogenesis and treatment of TMJOA.

Association between bilateral condylar resorption and reduced volumes of the craniofacial skeleton and masticatory muscles in adult patients: A retrospective study.

Objectives: This retrospective cohort study aimed to analyze volumes of craniomaxillofacial bone and masticatory muscles of young adults with bilateral idiopathic condylar resorption. Methods: This was a retrospective cohort study of 84 adults with bilateral idiopathic condylar resorption (BCR) and 48 adults with normal temporal-mandibular joint (TMJ) matched for age and sex (mean age, 23.2 ± 3.6 years). The volumes of craniomaxillofacial bone and masticatory muscles, as well as intercondylar angle were measured. Unpaired t-tests and Pearson correlation tests were applied to analyze the data. Multivariable linear regression models were used to estimate the association between bilateral condylar volume and volumes of craniomaxillofacial bone and masticatory muscles adjusted for age, sex, and disc status. Results: Compared to the control group, the BCR group displayed significant decreased volumes of craniomaxillofacial bone (p < 0.001), craniomaxillofacial bone without mandible (p < 0.001), mandible (p < 0.001), mandible without mandibular condylar process (p < 0.001), bilateral masseter muscle (p < 0.001) and bilateral temporalis muscle (p < 0.001), as well as the intercondylar angle (p < 0.001). These variables were significantly correlated to the volume of mandibular condylar process (0.5< r < 0.8; p < 0.001). By linear regression analyses, significant associations were found for the bilateral condylar volume with craniomaxillofacial bone volume and mandible bone volume. Conclusions: Young adults with BCR displayed smaller volumes of craniomaxillofacial skeleton and masticatory muscles, and smaller intercondylar angle than the normal patients. The craniofacial musculoskeletal volume and intercondylar angle are associated with mandibular condylar process volume.

Effect of arthroscopic discopexy on condylar growth in adolescents with temporomandibular joint disc displacement without reduction: A retrospective self-controlled case series study.

This study was a retrospective self-controlled study that aimed to evaluate the effect of arthroscopic discopexy on condylar height and mandibular position in adolescents with temporomandibular joint (TMJ) anterior disc displacement without reduction (ADDwoR). Patients between 10 and 20 years of age and diagnosed with bilateral TMJ ADDwoR by magnetic resonance image (MRI) were included in this study. All patients underwent a period of natural course before arthroscopic surgery and then a follow-up period postoperatively. Changes in condylar height and mandibular position were measured by MRI and X-ray radiographs. Data were analyzed by paired t-test, Pearson correlation analysis, and generalized estimating equations. This study comprised a total of 40 patients with a mean age of 14.80 years. Pearson correlation analysis showed correlations between condylar height and mandibular position changes. The condylar height change during the post-operative period was significantly higher than that during natural course period (3.57 mm, p < 0.001). The changes in mandibular position (including ANB angle, SNB angle, and Pog-Np) were significant different (all p < 0.05) between the two periods. This study found that arthroscopic discopexy can promote condylar growth and correct dentofacial deformity in adolescents with bilateral TMJ ADDwoR.

Single-cell RNA sequencing reveals neurovascular-osteochondral network crosstalk during temporomandibular joint osteoarthritis: Pilot study in a human condylar cartilage.

Purpose: Temporomandibular joint osteoarthritis (TMJ-OA) is one of the most complex temporomandibular disorders, causing pain and dysfunction. The main pathological feature of TMJ-OA is neurovascular invasion from the subchondral bone to the condylar cartilage. This study aimed to discover the cells and genes that play an important role in the neurovascular-osteochondral network crosstalk in human TMJ-OA. Materials and methods: Condylar cartilages from patient with TMJ-OA were divided into OA group, and others from patients with benign condylar hyperplasia (CH) were used as control for further single-cell RNA-sequencing (scRNA-seq). Hematoxylin and eosin staining were performed. The cells and genes in the condylar cartilage were identified and analyzed by scRNA-seq. Results: Histological analysis revealed blood vessel invasion and ossification in the TMJ-OA condylar cartilage. The scRNA-seq identified immune cells, endothelial cells, and chondrocytes in the TMJ-OA condylar cartilage. Macrophages, especially M1-like macrophages, contributed to the inflammation, angiogenesis, and innervation. CD31+ endothelial cells contributed to the bone mineralization. The TMJ-OA cartilage chondrocytes highly expressed genes related to inflammation, angiogenesis, innervation, and ossification. The hub genes contributing to these processes in the TMJ-OA chondrocytes included CTGF, FBN1, FN1, EGFR, and ITGA5. Conclusion: Our study marks the first time scRNA-seq was used to identify the cells and genes in a human TMJ-OA condylar cartilage, and neurovascular-osteochondral network crosstalk during the human TMJ-OA process was demonstrated. Targeting the crosstalk of these processes may be a potential comprehensive and effective therapeutic strategy for human TMJ-OA.

miR-335-5p inhibits endochondral ossification by directly targeting SP1 in TMJ OA.

OBJECTIVE: During the development of temporomandibular joint osteoarthritis, endochondral ossification is compromised which leads to condylar degeneration; miR-335-5p in endochondral ossification in osteoarthritic condylar cartilage tissue remains unclear. METHODS: Up-regulated microRNA and its target gene were searched for endochondral ossification in osteoarthritis articular cartilage. The effect of increased or decreased miR-335-5p on endochondral ossification was evaluated by transfecting miR-335-5p mimics or miR-335-5p inhibitor in vitro in chondrocytes C28/I2. Finally, we injected the temporomandibular joint of rats intra-articularly with agomiR-335 in a unilateral anterior crossbite rat model to determine the in vivo regulation of miR-335. RESULTS: After the onset of temporomandibular joint osteoarthritis, miR-335-5p levels were gradually up-regulated, whereas endochondral ossification-related genes were down-regulated in condylar cartilage specimens. Our results showed that miR-335 inhibited endochondral ossification after administration of a miR-335 antagonist into the temporomandibular joint articular cavity of a unilateral anterior crossbite rat model. AgomiR-335, a miR-335 agonist, inhibited matrix mineralization in fibrocartilage stem cells in vitro and then miR-335-5p negatively regulated chondrocyte activity by directly targeting SP1 via promoting transforming growth factor-ß/Smad signalling. CONCLUSION: miR-335-5p can significantly inhibit endochondral ossification; therefore, its inhibition may be beneficial for the treatment of temporomandibular joint osteoarthritis.

Chitosan based macromolecular hydrogel loaded total glycosides of paeony enhances diabetic wound healing by regulating oxidative stress microenvironment.

Oxidative stress microenvironment caused by reactive oxygen species (ROS) accumulation seriously hinders wound healing in diabetes, which brings great burden to global health. Various wound dressings on the market focus on delivering active substances to promote wound healing in diabetes. However, the complex pathological microenvironment of diabetic wounds often leads to the inactivation of delivery factors, which often leads to treatment failure, and thus, emerging therapeutic approaches are urgently needed. In this study, a macromolecular hydrogel synthesized by crosslinking N-carboxyethyl chitosan, hyaluronic acid-aldehyde, and adipic acid dihydrazide, with self-healing and injectable abilities was used to deliver total glycosides of paeony (TGP). The TGP incorporated hydrogel could obviously induce fibroblasts proliferation and secretion of various extracellular matrix proteins and growth factors, induce migration and angiogenesis of vein endothelial cells, and enhance macrophages polarization to M2 phenotype by eliminating accumulated ROS. In diabetic wound models, the ROS-scavenging hydrogel efficiently enhanced proliferation, re-epithelialization, collagen deposition, as well as angiogenesis in the wound area. Besides, the dressing induced the macrophages polarization from M1 phenotype (pro-inflammatory) to M2 phenotype (anti-inflammatory) and decreased the levels of inflammatory cytokines, thereby enhancing the diabetic wound healing. The wounds treated with TGP incorporated hydrogel almost completely healed 16 days after treatment. However, the residual wound areas in the groups of Con, INTRA, and Gel are 55.2 ± 4.6 %, 33.7 ± 6.5 %, and 34.9 ± 6.1 % on the 16th day, respectively. This hydrogel with pathological microenvironment improvement ability affords a novel therapeutic strategy for enhancing healing of chronic diabetic wound.

Corosolic acid-modified lipid nanoparticles as delivery carriers for DNA vaccines against avian influenza.

Cholesterol (CHOL) is essential for developing lipid nanoparticles (LNPs) for gene delivery because it enhances membrane fusion and improves the delivery efficiency of gene cargos. An attractive pDNA carrier, corosolic acid (CA)-modified lipid nanoparticles (CLNPs), was developed by replacing CHOL in LNPs to deliver pDNA at various ratios of nitrogen groups to phosphate groups (N/P). The resultant CLNPs with a higher CHOL/CA ratio exhibited similar mean particle size, zeta potential, and encapsulation efficiency to those of LNPs. In comparison with LNPs, CLNPs (CHOL:CA ratio = 2:1) achieved increased cellular uptake and enhanced transfection efficacy while maintaining low cytotoxicity. In vivo results from chicken experiments demonstrated that CLNPs encapsulating DNA vaccines against avian influenza at a N/P ratio of 3 could elicit similar-level humoral and cellular immune responses compared with those of LNPs at a higher N/P ratio, thereby suggesting the induction of desirable immune effects using less ionizable lipids. Our study provides a reference for further research on the application of CA in LNPs for gene delivery, and the development of novel delivery systems for DNA vaccines against avian influenza.

N-MID, P1NP, ß-CTX, and phosphorus in adolescents with condylar resorption.

OBJECTIVE: Condylar resorption (CR) is a temporomandibular joint disease that causes various physical or functional defects. We aimed to find the association between CR and bone metabolism levels. STUDY DESIGN: In this study, we recruited patients visiting the Orthodontic Clinic at Shanghai Ninth People's Hospital from January 2020 to September 2021. Patient characteristics, magnetic resonance imaging examination results, bone mineral density (BMD), Z-score, bone turnover markers, minerals, and hormones were collected and analyzed. RESULTS: The 89 participants were divided into CR (n = 46) and normal (n = 43) groups. Univariate logistic regression showed that N-terminal mid-fragment of osteocalcin (N-MID), procollagen type 1 N-terminal propeptide (P1NP), ß-C-terminal telopeptide of type 1 collagen (ß-CTX), and phosphorus (P < .001 for all) were protective factors, and BMD (P = .047) was a risk factor for CR. Multivariable logistic regression showed that N-MID, P1NP, ß-CTX, and phosphorus (odds ratio <1, P < .05 for all) were protective factors for CR. Receiver operating characteristic curves showed these indicators to effectively predict CR occurrence (area under the curve >0.7; P < .001). CONCLUSION: Adolescents with low serum N-MID, P1NP, ß-CTX, and phosphorus levels were associated with a higher risk of CR. We suggest that these indicators can guide clinicians in the early detection and prevention of CR in adolescents.

Rapid induction and long-term self-renewal of neural crest-derived ectodermal chondrogenic cells from hPSCs.

Articular cartilage is highly specific and has limited capacity for regeneration if damaged. Human pluripotent stem cells (hPSCs) have the potential to generate any cell type in the body. Here, we report the dual-phase induction of ectodermal chondrogenic cells (ECCs) from hPSCs through the neural crest (NC). ECCs were able to self-renew long-term (over numerous passages) in a cocktail of growth factors and small molecules. The cells stably expressed cranial neural crest-derived mandibular condylar cartilage markers, such as MSX1, FOXC1 and FOXC2. Compared with chondroprogenitors from iPSCs via the paraxial mesoderm, ECCs had single-cell transcriptome profiles similar to condylar chondrocytes. After the removal of the cocktail sustaining self-renewal, the cells stopped proliferating and differentiated into a homogenous chondrocyte population. Remarkably, after transplantation, this cell lineage was able to form cartilage-like structures resembling mandibular condylar cartilage in vivo. This finding provides a framework to generate self-renewing cranial chondrogenic progenitors, which could be useful for developing cell-based therapy for cranial cartilage injury.

Thermosensitive hydrogel loaded with concentrated growth factors promote bone repair in segmental bone defects.

Treating critical-size bone defects beyond the body's self-healing capacity is a challenging clinical task. In this study, we investigate the effect of concentrate growth factors (CGFs) loaded Poloxamer 407 hydrogel on the viability and osteogenic differentiation potential of bone marrow mesenchymal stem cells (BMSCs) and reconstruction of critical-size bone defects. In vitro, this CGFs-loaded thermosensitive hydrogel can significantly promote proliferation, maintain cell viability, and induce osteogenic differentiation of BMSCs by up-regulating the mineralization and alkaline phosphatase (ALP) activity, as well as gene markers, including runt-related transcription factor-2 (Runx-2), type I collagen (Col-1), osteocalcin (OCN), as well as osteopontin (OPN). In vivo, Micro-CT radiography analysis and histological detection demonstrated that the CGFs-loaded hydrogel significantly induced bone healing and reconstructed the medullary cavity structure in critical-size bone defect models. In conclusion, this strategy of transplantation of CGFs-loaded hydrogel promoted bone regeneration and prevented bone nonunion, so as to provide basis for clinical treatment for repairing critical-size bone defects.

Association between sexual maturation and anterior disc displacement of temporomandibular joint in adolescents aged 13-14 years.

OBJECTIVES: To investigate the association between sexual maturation and anterior disc displacement (ADD) of temporomandibular joint (TMJ). MATERIALS AND METHODS: Adolescents aged 13-14 years old, attending the first grade of one private junior school, were recruited. Magnetic resonance imaging (MRI) was used to confirm ADD, in addition, the serum levels of sex hormones were tested. Secondary sex characteristics, psychological evaluations, oral health-related behaviors, and sociodemographic characteristics were also collected. RESULTS: A total of 440 teenagers were included, of which the prevalence of ADD was 17.7%. Subgroup analysis revealed that age at menarche in girls and nocturnal emission, laryngeal prominence, voice change, and facial hair in boys showed significant differences between the ADD group and the normal population (all p < 0.05). The serum levels of prolactin in girls (12.23 and 9.82 ng/ml) and testosterone in boys (3.65 and 2.48 ng/dl) were significantly higher in the ADD group compared to the normal population, respectively (both p < 0.05). CONCLUSIONS: This study suggested that sexual maturation has a significant association with ADD both in boys and girls. The increased serum levels of testosterone in boys and prolactin in girls might contribute to the occurrence of TMJ ADD. CLINICAL RELEVANCE: This study provides key data to support and inform longitudinal studies as well as to explain why ADD prefers in adolescents. Main findings can also be used to prevent the occurrence of ADD and secondary jaw deformities.

Characteristics of patients with temporomandibular joint idiopathic condylar resorption.

OBJECTIVE: To investigate characteristics of temporomandibular joint (TMJ) idiopathic condylar resorption (ICR) and analyze the related factors. METHODS: A total of 755 consecutive patients (150 with ICR and 605 with anterior disc displacement [ADD]) from July 2015 to December 2018 were recruited. A comprehensive questionnaire characterizing the multidimensional impact of the TMJ was designed. Clinical examination and radiological evaluation were also performed. The odds ratio for each variable in the ICR group versus the ADD group was computed using logistic regression analysis. RESULTS: Multivariate logistic regression analysis showed significant correlations between mouth opening restriction, disease course, mandibular retrusion, mandibular retrusion progression, skeletal Class II profile, and overjet in ICR patients. CONCLUSION: These results suggest that a longer ADD disease course might have a strong relationship with ICR.

UCHL1 promotes proliferation and metastasis in head and neck squamous cell carcinoma and could be a potential therapeutic target.

OBJECTIVE: The purpose of this study was to research the physiological roles of ubiquitin carboxyl-terminal esterase L1 (UCHL1) in head and neck squamous cell carcinoma (HNSCC). STUDY DESIGN: Ten HNSCC samples and matched normal oral mucosal tissues were collected. UCHL1 expression of these tissues was detected by the immunohistochemical staining and real-time quantitative polymerase chain reaction. The human HNSCC cell line HN6 UCHL1 knockout (UCHL1 KO) cell line was constructed using CRISPR/CAS9 gene editing and verified by western blotting. Wound healing assay, cell proliferation assay, cell invasion assay, and flow cytometric analysis of the cell cycle and apoptosis were applied to research the role of UCHL1 in HNSCC. Also, an RNAseq gene expression data set and HNSCC patient survival data from The Cancer Genome Atlas were analyzed. RESULTS: UCHL1 was highly expressed in HNSCC tissues compared with normal oral mucosal tissues (P = .032). A decreased proliferation (P < .0001), migration (P < .0001), and invasion (P = .0049) ability of HN6 cells was exhibited after knockout of UCHL1. However, HN6 UCHL1 KO cells showed no significant differences in the cell cycle or apoptosis. The progression, nodal metastasis status, and stage of HNSCC had a positive correlation with the expression of UCHL1. CONCLUSIONS: UCHL1 plays an important role in HNSCC, and we consider that targeting UCHL1 may be a feasible therapeutic strategy for HNSCC.

Identification of four key prognostic genes and three potential drugs in human papillomavirus negative head and neck squamous cell carcinoma.

BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) is a common tumor worldwide with poor prognosis. The pathogenesis of human papillomavirus (HPV)-positive and HPV-negative HNSCCs differs. However, few studies have considered the HPV status when identifying biomarkers for HNSCC. Thus, the identification of biomarkers for HPV-positive and HPV-negative HNSCCs is urgently needed. METHODS: Three microarray datasets from Gene Expression Omnibus (GEO) were analyzed, and the differentially expressed genes (DEGs) were obtained. Then, functional enrichment pathway analysis was performed and protein-protein interaction (PPI) networks were constructed. The expression of hub genes at both the mRNA and protein level was determined in Oncomine, The Cancer Genome Atlas (TCGA) and the Human Protein Atlas (HPA). In addition, survival analysis of the patient stratified by HPV status and the expression levels of key genes were performed based on TCGA data. The role of AREG, STAG3, CAV1 and C19orf57 in cancer were analyzed through Gene set enrichment analysis (GSEA). The top ten small molecule drugs were identified and the therapeutic value of zonisamide, NVP-AUY922, PP-2 and fostamatinib was further evaluated in six HPV-negative HNSCC cell lines. Finally, the therapeutic value of NVP-AUY922 was tested in vivo based on three HPV-negative HNSCC models, and statistical analysis was performed. RESULTS: In total, 47 DEGs were obtained, 11 of which were identified as hub genes. Biological process analysis indicated that the hub genes were associated with the G1/S transition of the mitotic cell cycle. Survival analysis uncovered that the prognostic value of AREG, STAG3, C19orf57 and CAV1 differed between HPV-positive and HPV-negative patients. Gene set enrichment analysis (GSEA) showed the role of AREG, STAG3 and CAV1 in dysregulated pathways of tumor. Ten small molecules were identified as potential drugs specifically for HPV-positive or HPV-negative patients; three-NVP-AUY922, fostamatinib and PP-2-greatly inhibited the proliferation of six HPV-negative HNSCC cell lines in vitro, and NVP-AUY922 inhibited three HPV-negative HNSCC xenografts in vivo. CONCLUSIONS: In conclusion, AREG, STAG3, C19orf57 and CAV1 are key prognostic factors and potential therapeutic targets in HPV-negative HNSCC. NVP-AUY922, fostamatinib and PP-2 may be effective drugs for HPV-negative HNSCC.

Long Non-Coding RNA Expression Profile Associated with Malignant Progression of Oral Submucous Fibrosis.

Oral submucous fibrosis (OSF) as one of the premalignant disorders endures a series of histopathological stages to invasive oral squamous cell carcinoma (OSCC) eventually. However, the role of long non-coding RNA (lncRNA) expression in OSF malignant progression still remains poorly understood. Through RNA-sequencing normal mucous, OSF and OSCC tissues, we found 687 lncRNA transcripts significantly and differentially expressed during OSF progression, including 231 upregulated lncRNAs and 456 downregulated lncRNAs, indicating that lncRNAs are involved in the regulation of different stages of OSF development. Further functional enrichment analysis showed these differentially expressed lncRNAs participated in inflammation signaling, Wnt signaling, angiogenesis, CCKR signaling, integrin signaling, PDGF signaling, p53 signaling, and EGF receptor (EGFR) signaling pathways, which contribute to inflammatory and fibroelastic pathogenetic changes of OSF and further malignant progression. Five novel lncRNAs were differentially expressed during OSF progression with varied expression levels, indicating the importance of these lncRNAs in OSF malignant development. Moreover, some lncRNAs have been previously identified to be associated with OSCC pathogenesis, including HCG22, RP11-397A16.1, LINC00271, CTD-3179P9.1, and ZNF667-AS1. Thus, our study firstly comprehensively elucidated lncRNAs expression profile of malignant procession from OSF premalignant lesion to OSCC, which will enlighten our understanding of the importance of lncRNA involved in OSF malignant development.

Hydrodynamic investigation of a novel shear-generating device for the measurement of anchorage-dependent cell adhesion intensity.

Proliferation of anchorage-dependent cells occurs after adhesion to a suitable surface. Thus, quantitative information about the force of cells adhesion to microcarriers at early culture phases is vital for scaling up such system. In this work, a newly designed shear-generating device was proposed, based on a previously proposed contraction flow device designed for suspended cells. A design equation for the new device was also proposed to correlate the generated energy dissipation rate (EDR) with the cross-sectional area and flow rate. Microscopic-particle image velocimetry was measured to validate the simulation results, and good agreement was achieved. The newly designed device was then used to measure the adhesion force of MDCK and PK cells, and the results showed that the critical EDR was 3000 W/m3 for MDCK and 5000 W/m3 for PK cells. This quantitative information is of great value for better understanding shearing effects during the scaling up of anchorage-dependent cell cultures.