Enhanced VEGF-A expression and mediated angiogenic differentiation in human gingival fibroblasts by stimulating with TNF-α in vitro.

Backgroud/purpose: The effects of inflammatory cytokines were reported to involve in the process of periodontal disease and inflamed tissue enhanced the expression of inflammatory mediators which in turn may promote angiogenesis. Human gingival fibroblasts (HGFs) exert the basic function in periodontal tissue repair and regeneration. However, studies specially focused on the effects of inflammation-related HGFs on angiogenic and osteogenic differentiation are limited. This study was aimed to test whether HGFs enhance vascular endothelial growth factor (VEGF)-A expression mediating angiogenic and osteogenic differentiation by stimulating with tumor necrosis factor-α (TNF-α), to further identify the possible mechanism which may be responsible for this activity. Materials and methods: In this study, HGFs are treated by TNF-α in order to detect the effects of angiogenic and osteogenic differentiation under inflammation-related condition. Results: TNF-α enhances VEGF-A expression and results in increasing cell migration and angiogenic differentiation and inhibiting osteogenic differentiation in HGFs. Besides, TNF-α stimulated VEGF-A-mediated angiogenic differentiation is dependent on the activation of mitogen-activated protein kinase (MAPK) pathway, Extracellular signal-regulated kinase (ERK) 1/2 phosphorylation may contribute to regulate the function of VEGF-A in inflammation-related HGFs. Conclusion: This study demonstrated that enhanced VEGF-A-mediated angiogenic differentiation in HGFs is dependent on the activation of MAPK pathway by stimulating with TNF-α in vitro. Therefore, this study could provide better understand for the progression of inflammation-related periodontal diseases.

Effect of diode laser combined with minocycline hydrochloride in nonsurgical periodontal therapy: a randomized clinical trial.

BACKGROUND: This study aimed to assess the effect of diode laser combined with minocycline hydrochloride in conventional nonsurgical periodontal therapy. METHODS: Ninety-two patients and 1206 teeth were included in this study. The patients were diagnosed moderate or severe periodontal diseases with the presence of teeth in at least 3 quadrants in the oral cavity. Each patient's quadrants were randomly divided into three treatment groups as following, Control group: scaling and root planning (SRP); Experimental group 1 (Exp 1): SRP + minocycline hydrochloride; Experimental group 2 (Exp 2): SRP + 809 nm diode laser + minocycline hydrochloride. The minocycline in Exp 1 and Exp 2 was applied once per week, for 4 weeks. Clinical examinations including periodontal probing depth (PD), clinical attachment level (CAL) and bleeding index (BI), and the secretion of inflammatory factor (tumor necrosis factor, TNF-α) was detected by ELISA before and 3, 6 months after the treatments. The differences among these groups were assessed by One-Way ANOVA and Kruskal-Wallis test. P-value < 0.05 was considered significant. RESULTS: All the periodontal indexes (PD, CAL and BI) were improved after each treatment and the secretion of TNF-α was reduced for all three groups. In patients with deep periodontal pockets, Exp 2 showed significant improvements in all indexes comparison with Con group and Exp group 1. CONCLUSIONS: The synergistic effect of SRP and 809 nm diode laser combined with minocycline hydrochloride could play an efficient and reliable effect in the nonsurgical periodontal treatment approach. Trial registration The clinical trial was retrospectively registered in chictr.org.cn with registration ChiCTR2100051708 (01/10/2021).

The miR-193a-3p-MAP3k3 Signaling Axis Regulates Substrate Topography-Induced Osteogenesis of Bone Marrow Stem Cells.

Substrate topographical features induce osteogenic differentiation of bone marrow stem cells (BMSCs), but the underlying mechanisms are unclear. As microRNAs (miRNAs) play key roles in osteogenesis and bone regeneration, it would be meaningful to elucidate the roles of miRNAs in the intracellular signaling cascade of topographical cue-induced osteogenic differentiation. In this study, the miRNA expression profile of the topographical feature-induced osteogenic differentiation group is different from that of the chemical-factors-induced osteogenic differentiation group. miR-193a-3p is sensitive to substrate topographical features and its downregulation enhances osteogenic differentiation only in the absence of osteogenesis-inducing medium. Also, substrate topographical features specifically activate a nonclassical osteogenetic pathway-the mitogen-activated protein kinase (MAPK) pathway. Loss- and gain-of-function experiments demonstrate that miR-193a-3p regulates the MAPK pathway by targeting the MAP3k3 gene. In conclusion, this data indicates that different osteogenic-lineage-related intracellular signaling cascades are triggered in BMSCs subjected to biophysical or chemical stimulation. Moreover, the miR-193a-3p-MAP3k3 signaling axis plays a pivotal role in the transduction of biophysical cues from the substrate to regulate the osteogenic lineage specification of BMSCs, and hence may be a promising molecular target for bone regenerative therapies.

An injectable double-network hydrogel for the co-culture of vascular endothelial cells and bone marrow mesenchymal stem cells for simultaneously enhancing vascularization and osteogenesis.

The achievement of rapid vascularization in large implanted constructs is a major challenge in the field of bone tissue engineering. Although co-culture of bone-forming cells and vascular endothelial cells (VECs) has been expected to be a way of promoting vascularization during bone formation with a scaffold, there is a lack of detailed knowledge about the direct interactions between two types of stem cells in a three-dimensional (3D) extracellular matrix (ECM). Herein, we report on the use of an injectable cytocompatible double-network (DN) hydrogel to encapsulate, co-culture and subsequently stimulate the angiogenic/osteogenic differentiation of VECs and the human bone marrow mesenchymal stem cells (BM-MSCs), which demonstrates that the direct co-cultured system enables simultaneous enhancement of vascularization and osteogenesis by providing 3D cell-cell communication. Besides, the improved mechanical properties and the injectability of the DN hydrogel allow the delivery, long-time implantation, proliferation and differentiation of stem cells in vivo. Therefore, this study could provide a niche-like native ECM for stem cell survival and the regulation of the differentiation of multiple cell lines which will benefit bone repair.

Tumor-associated macrophages correlate with the clinicopathological features and poor outcomes via inducing epithelial to mesenchymal transition in oral squamous cell carcinoma.

BACKGROUND: Both tumor-associated macrophages (TAMs) and the epithelial to mesenchymal transition (EMT) of cancer cells play key roles in promoting tumor progression. However, whether TAMs could induce EMT in the progression of oral squamous cell carcinoma (OSCC) remains undefined. RESULTS: Here we detected the expression of macrophages markers CD68 and CD163, epithelial marker E-cadherin and mesenchymal marker vimentin in 127 OSCC patients by using semi-quantitative immunohistochemistry. CD68 and CD163 expression was not confined to the infiltrating TAMs, but also detected in cancer cells. The high number of CD68-positive macrophages was correlated with poor overall survival. Meanwhile, the expression of CD163 both in macrophages and in cancer cells was associated with poor overall survival and had a significant prognostic impact in OSCC. Importantly, the expression of CD163 in cancer cells had a significant relationship with E-cadherin and vimentin. Furthermore, the incubation of TAMs conditioned medium resulted in a fibroblast-like appearance of cancer cells (HN4, HN6 and SCC9) together with the decreased/increased expression of E-cadherin/ vimentin, which were correlated with the enhanced ability of migration and invasion. CONCLUSIONS: Our results indicate that TAMs could promote the EMT of cancer cells, thereby leading to the progression of oral cancer.

Snail interacts with Id2 in the regulation of TNF-α-induced cancer cell invasion and migration in OSCC.

The inflammatory tumor microenvironment has been identified to play a pivotal role in tumor development and metastasis. Tumor necrosis factor-α (TNF-α) is one of the key cytokines that regulate the inflammatory processes in tumor promotion. In the current study, we treated three oral squamous cell carcinoma (OSCC) cell lines with TNF-α to study its role in inflammation-induced tumor progression. Here we show that TNF-α induces stabilization of the transcriptional repressor Snail and activates NF-κB pathway in the three OSCC cell lines. These activities resulted in the increased motility and invasiveness of three OSCC cell lines. In addition, upon dealing with TNF-α for the indicated time, three OSCC cell lines underwent epithelial-to-mesenchymal transition (EMT), in which they presented a fibroblast-like phenotype and had a decreased expression of epithelial marker (E-cadherin) and an increased expression of mesenchymal marker (vimentin). We further demonstrated that TNF-α can up-regulate the expression of Id2 while inducing an EMT in oral cancer cells. Finally, we showed that Id2 interacted with Snail which may constrain Snail-dependent suppression of E-cadherin. In conclusion, our study indicates that TNF-α induces Snail stabilization is dependent on the activation of NF-κB pathway and results in increasing cell invasion and migration in OSCC cells. Id2 may contribute to regulate the function of Snail during TNF-α-mediated EMT in OSCC. These findings have significant implications for inflammation-induced tumor promotion in OSCC.

Membrane Type 1 Matrix Metalloproteinase induces an epithelial to mesenchymal transition and cancer stem cell-like properties in SCC9 cells.

BACKGROUND: Tissue invasion and metastasis are acquired abilities of cancer and related to the death in oral squamous cell carcinoma (OSCC). Emerging observations indicate that the epithelial-to-mesenchymal transition (EMT) is associated with tumor progression and the generation of cells with cancer stem cells (CSCs) properties. Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) is a cell surface proteinase, which is involved in degrading extracellular matrix components that can promote tumor invasion and cell migration. METHODS: In the current study, we utilized SCC9 cells stably transfected with an empty vector (SCC9-N) or a vector encoding human MT1-MMP (SCC9-M) to study the role of MT1-MMP in EMT development. RESULTS: Upon up-regulation of MT1-MMP, SCC9-M cells underwent EMT, in which they presented a fibroblast-like phenotype and had a decreased expression of epithelial markers (E-cadherin, cytokeratin18 and ß-catenin) and an increased expression of mesenchymal markers (vimentin and fibronectin). We further demonstrated that MT1-MMP-induced morphologic changes increased the level of Twist and ZEB, and were dependent on repressing the transcription of E-cadherin. These activities resulted in low adhesive, high invasive abilities of the SCC9-M cells. Furthermore, MT1-MMP-induced transformed cells exhibited cancer stem cell (CSC)-like characteristics, such as low proliferation, self-renewal ability, resistance to chemotherapeutic drugs and apoptosis, and expression of CSCs surface markers. CONCLUSIONS: In conclusion, our study indicates that overexpression of MT1-MMP induces EMT and results in the acquisition of CSC-like properties in SCC9 cells. Our growing understanding of the mechanism regulating EMT may provide new targets against invasion and metastasis in OSCC.

Snail overexpression induces an epithelial to mesenchymal transition and cancer stem cell-like properties in SCC9 cells.

Local invasiveness and distant metastasis are critical factors that contribute to oral squamous cell carcinoma-related deaths. Increasing evidence has shown that the epithelial to mesenchymal transition (EMT) is involved in cancer progression and is associated with the 'stemness' of cancer cells. Snail is a transcriptional factor that can induce EMT and preserve stem-cell function, which may induce resistance to radio- and chemotherapies in the cells. In the present study, SCC9 cells were transfected with an empty vector or a vector encoding human Snail (SCC9-S). Overexpression of Snail induced SCC9 cells to undergo EMT, in which the cells presented a fibroblast-like appearance, downregulated the epithelial markers E-cadherin and ß-catenin, upregulated the mesenchymal marker vimentin, and associated with highly invasive and metastatic properties. Furthermore, the induction of EMT promoted cancer stem cell (CSC)-like characteristics in the SCC9-S cells, such as low proliferation, self-renewal, and CSC-like markers expression. These results indicate that overexpression of Snail induces EMT and promotes CSC-like traits in the SCC9 cells. Further understanding the role of Snail in cancer progression may reveal new targets for the prevention or therapy of oral cancers.