Differential expression of circular RNAs in interleukin 6-promoted osteogenic differentiation of human stem cells from apical papilla.

INTRODUCTION: Studies have shown that interleukin 6 (IL-6) can regulate stem cell osteogenic differentiation; however, the exact mechanism is not clear. Circular RNAs (circRNAs) are closed circular non-coding RNAs that are involved in the process of stem cell osteogenic differentiation. Therefore, the purpose of this present study was to investigate the effect of IL-6 treatment on osteogenic differentiation of human apical tooth papillae stem cells (hSCAPs), and to detect the difference in circRNA expression using gene microarray technology. METHODS: After extraction and identification of hSCAPs, alkaline phosphatase (ALP) activity, alizarin red staining, and calcium ion quantitative assay were used to determine the changes of ALP enzyme, mineralized nodules, and matrix calcium levels before and after IL-6 treatment of hSCAPs gene microarray technology was used to analyze the changes in circRNA expression levels before and after IL-6 induction of mineralization. The four selected circRNAs were validated by qRT-PCR. Moreover, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to predict the potential functions and biological signaling pathways of circRNAs. Finally, these data are integrated and analyzed to construct circRNA-microRNA-mRNA networks. RESULTS: Alp and Alizarin red staining confirmed that IL-6 promoted the osteogenic differentiation of hSCAPs. The gene microarray results identified 132 differentially expressed circRNAs, of which 117 were upregulated and 15 were downregulated. Bioinformatic analysis predicted that the circRNA-406620/miR-103a-3p/FAT atypical cadherin 4 (FAT4) pathway might be involved in regulating IL-6 to promote osteogenic differentiation of hSCAPs. CONCLUSION: Differentially expressed circRNAs might be closely involved in regulating IL-6 to promote osteogenic differentiation of hSCAPs.

miR-221-3p and miR-222-3p downregulation promoted osteogenic differentiation of bone marrow mesenchyme stem cells through IGF-1/ERK pathway under high glucose condition.

OBJECTIVE: This study aims to investigate the roles of miR-221-3p and miR-222-3p in the regulation of osteogenic differentiation of bone marrow mesenchyme stem cells (BMSCs) under high glucose condition. MATERIALS AND METHODS: The expreesions of miR-221-3p, miR-222-3p and insulin-like growth factor 1 (IGF-1) were detected by qRT-PCR. The protein levels of osteoblast-related proteins (Osterix, Runx-2 and Osteopontin) were detected by western blot. Whether miR-221-3p and miR-222-3p can target IGF-1 was assessed by dual luciferase reporter gene assay. RESULTS: miR-221-3p and miR-222-3p were up-regulated in the mandibles of diabetic rats and BMSCs cultured in high glucose condition. Silencing miR-221-3p or/ and miR-222-3p increased ALP activity and up-regulated osteoblast-related protein levels, and the simultaneous silence the two miRNAs showed stronger effects on ALP activity and osteoblast-related protein levels. Next, we confirmed that miR-221-3p and miR-222-3p both targeted IGF-1 and cooperatively regulated its expression. Besides, miR-221-3p and miR-222-3p regulated ERK activation through IGF-1. Silencing miR-221-3p and miR-222-3p promoted osteogenic differentiation of BMSCs through IGF-1 under high glucose condition. CONCLUSION: miR-221-3p and miR-222-3p inhibited osteogenic differentiation of BMSCs via IGF-1/ERK pathway under high glucose condition.

Significance of the neutrophil-to-lymphocyte ratio in young patients with oral squamous cell carcinoma.

BACKGROUND: The main goal of this study was to evaluate the prognosis of young patients with oral squamous cell carcinoma (SCC) with a focus on the value of the pretreatment neutrophil-to-lymphocyte ratio (NLR). MATERIALS AND METHODS: Young (≤40 years old) patients with oral SCC were retrospectively enrolled, and each young patient was matched with an old (≥60 years old) oral SCC patient. Associations between the NLR and clinicopathological variables were analyzed by the chi-square test, and the Kaplan-Meier method was used to analyze recurrence-free survival (RFS) and disease-specific survival (DSS) rates. RESULTS: A total of 103 young patients were enrolled, and compared to the old group, the young group had a significantly lower NLR value (p=0.012). In the young group, the 5-year RFS and DSS rates were 82% and 85%, respectively. In the old group, the 5-year RFS and DSS rates were 65% and 71%, respectively, and the differences between the groups were significant (both p<0.05). In the young patients with an NLR≤2.56, the 5-year DSS rate was 93%, while in the young patients with an NLR >2.56, the 5-year DSS rate was 76%. This difference was significant (p=0.020). A further Cox model analysis confirmed that the NLR was an independent prognostic factor for DSS. CONCLUSION: Young patients with oral SCC have a better prognosis than old oral SCC patients, and the NLR is significantly associated with DSS in young patients.

Preparation and characterization of general-purpose gelatin-based co-loading flavonoids nano-core structure.

Flavonoids (FLAs) possess anti-cancer, anti-viral, anti-bacterial, and anti-oxidant properties. In this study, gelatin nanoparticles (GNPs) with controllable surface potential and diameter was prepared through a modified two-step desolvation. Two well-known flavonoids, namely, low-molecular weight Genistein (GEN) and high-molecular weight Icariin (ICA), were adsorbed onto the surface of GNPs (FLA@GNPs). The characteristics of GNPs and the main parameters affecting flavonoid adsorption were studied to evaluate the adsorption capacity and structural stability of FLA@GNPs. Furthermore, co-adsorption of GEN and ICA was detected. The adsorption mechanism of GNPs with FLA was further discussed. Results showed that the low-molecular weight GEN could be effectively adsorbed by GNPs, and their entrapment efficiencies were over 90% under optimized conditions. The total drug loading of the co-adsorbed FLA@GNPs was significantly higher than that of the single drug loaded (GEN or ICA). GEN@GNPs could maintain its structural stability under acidic conditions (pH = 2) at room temperature (25 °C). This protective function enables both ICA and GEN to be bioactive at room temperature for at least 180 days. The characteristics of GNPs adsorption indicate that the hydrogen bonding theory of the combination of gelatin molecules with polyphenols cannot sufficiently explain the binding of GNPs with polyphenols. FLA@GNPs is a promising general-purpose gelatin-based co-loading preload structure with simplified operation and storage condition.