Analysis of ceRNA networks during mechanical tension-induced osteogenic differentiation of periodontal ligament stem cells.

The molecular mechanisms underlying osteogenic differentiation of periodontal ligament stem cells (PDLSCs) under mechanical tension remain unclear. This study aimed to identify a potential long non-coding ribonucleic acids (lncRNAs)/circular RNAs (circRNAs)-microRNAs (miRNAs)-messenger RNAs (mRNAs) network in mechanical tension-induced osteogenic differentiation of PDLSCs. PDLSCs were isolated from the healthy human periodontal ligament, identified, cultured, and exposed to tensile force. The expression of osteogenic markers was examined, and whole transcriptome sequencing was performed to identify the expression patterns of lncRNA, circRNA, miRNAs, and mRNAs. Enrichment analyses were also performed. Candidate targets of differentially expressed non-coding RNAs (ncRNAs) were predicted, and potential competitive endogenous RNA (ceRNA) networks were constructed by Cytoscape. We found that the osteogenic differentiation of PDLSCs was significantly enhanced under dynamic tension (magnitude: 12%, frequency: 0.7 Hz). Overall, 344 lncRNAs, 57 miRNAs, 41 circRNAs, and 70 mRNAs were differentially expressed in the tension group and the control group. Functional enrichment analysis showed that differentially expressed mRNAs were mainly enriched in osteogenesis-related and mechanical stress-related biological processes and signal transduction pathways (e.g., tumor necrosis factor [TNF] and Hippo signaling pathways). The lncRNA/circRNA-miRNA-mRNA networks were depicted, and potential key ceRNA networks were identified. Our findings may help to further explore the underlying regulatory mechanism of osteogenic differentiation of PDLSCs under mechanical tensile stress.

Wang W ，Li N ，Wang M ，Zhao Y ，Wu H ，Shi J ，Musa M ，Chen X ... -  《-》

Integrative Analysis of ceRNA Networks in human periodontal ligament stem cells under hypoxia.

This study aims to investigate the regulatory effect of hypoxia on human periodontal ligament stem cells (PDLSCs) through RNA sequencing (RNA SEQ). Human PDLSCs were cultured in normoxia (20% O2 ) or hypoxia (2% O2 ). Total RNA was extracted and sequenced. The expression profiles of circRNAs, lncRNAs, and miRNAs were determined, and the lncRNA/circRNA-miRNA-mRNA networks were analyzed. In total, 15 miRNAs, 449 lncRNAs, and 53 circRNAs were differentially expressed. Among them, 21 circRNAs, 262 lncRNAs, 5 miRNAs, and 5 mRNAs were selected to construct competing endogenous RNA (ceRNA) networks. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were carried out to explore potential related pathways and regulatory functions. Several ceRNA axes (lncRNA-FTX/circRNA-FAT1-hsa-miR-4781-3p-SMAD5 and circRNA LPAR1-hsa-miR-342-3p-ADAR) may provide a theoretical basis on the study of osteogenic differentiation of PDLSCs under hypoxia. This study revealed that the expression profiles of circRNAs, lncRNAs, and miRNAs had changed significantly in PDLSCs cultured in 2% O2 ; specific circRNAs/lncRNAs may play a competitive role in the differentiation of PDLSCs.

Ye Y ，Fu L ，Liu L ，Xiao T ，Cuba Manduca AG ，Yu J ... -  《-》

Identification and integrated analysis of differentially expressed lncRNAs and circRNAs reveal the potential ceRNA networks during PDLSC osteogenic differentiation.

Gu X ，Li M ，Jin Y ，Liu D ，Wei F ... -  《BMC GENETICS》

Analysis of lncRNAs-miRNAs-mRNAs networks in periodontal ligament stem cells under mechanical force.

Our study aims to analyze the expression profiles of long non-coding RNAs (lncRNAs) and investigate the potential regulatory networks among lncRNAs, microRNAs (miRNAs), and mRNAs in periodontal ligament stem cells (PDLSCs) under mechanical force (MF). PDLSCs were isolated from human periodontal ligament tissues and identified by flow cytometry analysis. Multidirectional differentiation potential of PDLSCs was obtained by osteogenic and adipogenic induction. High-throughput RNA sequencing was used to identify the expression patterns of lncRNAs and mRNAs in PDLSCs under MF. MF-responsive miRNAs were obtained from the previous microarray data. LncRNAs-miRNAs-mRNAs networks were constructed by Cytoscape. PDLSCs cultured from the periodontal ligament tissues were positive for STRO-1, CD146 and negative for CD45, CD34. Alizarin red staining and Oil Red O staining showed that PDLSCs had the ability of osteogenic and adipogenic differentiation. Then, a total of 1,339 and 1,426 differentially expressed lncRNAs and mRNAs were identified, respectively, in PDLSCs under MF. Based on the previous miRNA microarray analysis, the potential interaction networks of lncRNAs-miRNAs-mRNAs were constructed. It was found that lncRNAs and mRNAs could competitively interact with the same miRNA. LncRNAs-miRNAs-mRNAs networks were involved in PDLSCs under MF, which might provide a novel mechanism in the regulation of clinical orthodontic tooth movement process.

Wang H ，Feng C ，Li M ，Zhang Z ，Liu J ，Wei F ... -  《-》

Comprehensive analysis of the long noncoding RNA-associated competitive endogenous RNA network in the osteogenic differentiation of periodontal ligament stem cells.

The mechanism implicated in the osteogenesis of human periodontal ligament stem cells (PDLSCs) has been investigated for years. Previous genomics data analyses showed that long noncoding RNA (lncRNA), microRNA (miRNA) and messenger RNA (mRNA) have significant expression differences between induced and control human PDLSCs. Competing for endogenous RNAs (ceRNA), as a widely studied mechanism in regenerative medicine, while rarely reported in periodontal regeneration. The key lncRNAs and their ceRNA network might provide new insights into molecular therapies of periodontal regeneration based on PDLSCs. Two networks reflecting the relationships among differentially expressed RNAs were constructed. One ceRNA network was composed of 6 upregulated lncRNAs, 280 upregulated mRNAs, and 18 downregulated miRNAs. The other network contained 33 downregulated lncRNAs, 73 downregulated mRNAs, and 5 upregulated miRNAs. Functional analysis revealed that 38 GO terms and 8 pathways related with osteogenesis were enriched. Twenty-four osteogenesis-related gene-centred lncRNA-associated ceRNA networks were successfully constructed. Among these pathways, we highlighted MAPK and TGF-beta pathways that are closely related to osteogenesis. Subsequently, subnetworks potentially linking the GO:0001649 (osteoblast differentiation), MAPK and TGF-beta pathways were constructed. The qRT-PCR validation results were consistent with the microarray analysis. We construct a comprehensively identified lncRNA-associated ceRNA network might be involved in the osteogenesis of PDLSCs, which could provide insights into the regulatory mechanisms and treatment targets of periodontal regeneration.

Lai L ，Wang Z ，Ge Y ，Qiu W ，Wu B ，Fang F ，Xu H ，Chen Z ... -  《-》