circRNA Expression Profile in Dental Pulp Stem Cells during Odontogenic Differentiation

Abstract

Introduction. Odontogenic differentiation of human dental pulp stem cells (hDPSCs) is a key step of pulp regeneration. Recent studies showed that circular RNAs (circRNAs) have many biological functions and that competing endogenous RNA (ceRNA) is their most common mechanism of action. However, the role of circRNAs in hDPSCs during odontogenesis is still unclear. Methods. Isolated hDPSCs were cultured in essential and odontogenic medium. Total RNA was extracted after 14 days of culture, and then, microarray analysis was performed to measure the differential expressions of circRNAs. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was then performed to validate the microarray results. Based on microarray data from this study and available in the database, a ceRNA network was constructed to investigate the potential function of circRNAs during odontogenesis. In addition, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to investigate the potential correlation between signaling pathways and circRNAs. In addition, qRT-PCR and Western blot analysis were used to explore the function of hsa_circRNA_104101. Results. We found 43 upregulated circRNAs and 144 downregulated circRNAs during the odontogenic differentiation process (fold change>1.5 and <-1.5, respectively; P<0.05). qRT-PCR results were in agreement with the microarray results. Bioinformatic analysis revealed that the Wnt signaling pathway and the TGF-β signaling pathway, as well as the other pathways associated with odontogenic differentiation, were correlated to the differentially expressed circRNAs. hsa_circRNA_104101 was proved to promote the odontogenic differentiation of hDPSCs. Conclusion. This study reported 187 circRNAs that were differentially expressed in hDPSCs during odontogenic differentiation. Bioinformatic analysis of the expression data suggested that circRNA-miRNA-mRNA networks might act as a crucial mechanism for hDPSC odontogenic differentiation, providing a theoretical foundation for the study of pulp regeneration regulation by circRNAs.

The Emerging Regulatory Role of Circular RNAs in Periodontal Tissues and Cells

“…These studies have focussed on the role of circRNA in controlling cell differentiation, which is important in the context of periodontal regeneration and tissue engineering. Herein, we undertook a scoping review of recent research into the role of circRNA in cell differentiation in vitro and periodontal tissue healing and regeneration in vivo, drawing information from 23 studies [ 37 , 38 , 39 , 40 , 41 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 ]. Much of the existing research has focused on osteogenic differentiation of periodontal stem cells, and how circRNAs might function as a sponge for miRNAs.…”

N-Cadherin Regulates the Odontogenic Differentiation of Dental Pulp Stem Cells via β-Catenin Activity

“…Healthy dental pulp tissues were collected from caries-free teeth of patients (age 18–25 years old) undergoing extraction of fully erupted third molars, according to the informed protocol approved by the Ethics Committee of Nanfang Hospital, Southern Medical University, Guangzhou, China. Primary human DPSCs were isolated, identified and cultured in Dulbecco’s modified Eagle’s medium (Gibco, Grand Island, NY, United States) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco), as described previously ( Chen et al, 2020 ). The cells were maintained at 37°C in a humidified atmosphere with 5% CO 2 and 95% air.…”

Circular RNAs in stem cell differentiation: a sponge-like role for miRNAs

“…A microarray analysis revealed 43 upregulated circRNAs and 144 downregulated circRNAs to be differentially expressed in human DPSCs during odontogenic differentiation process. Moreover, hsa_circRNA_104101 was highly assumed to enhance the odontogenic differentiation of human DPSCs 37 . Based on above, it is well established that the common mechanisms of circRNAs in DPSCs are competently binding with miRNAs to deliver a promoting effect on osteogenic differentiation ( Figure 2 A ).…”