IGF-1/IGF-1R/hsa-let-7c axis regulates the committed differentiation of stem cells from apical papilla

Abstract

Insulin-like growth factor-1 (IGF-1) and its receptor IGF-1R play a paramount role in tooth/bone formation while hsa-let-7c actively participates in the osteogenic differentiation of mesenchymal stem cells. However, the interaction between IGF-1/IGF-1R and hsa-let-7c on the committed differentiation of stem cells from apical papilla (SCAPs) remains unclear. In this study, human SCAPs were isolated and treated with IGF-1 and hsa-let-7c over/low-expression viruses. The odonto/osteogenic differentiation of these stem cells and the involvement of mitogen-activated protein kinase (MAPK) pathway were subsequently investigated. Alizarin red staining showed that hsa-let-7c low-expression can significantly promote the mineralization of IGF-1 treated SCAPs, while hsa-let-7c over-expression can decrease the calcium deposition of IGF-1 treated SCAPs. Western blot assay and real-time reverse transcription polymerase chain reaction further demonstrated that the expression of odonto/osteogenic markers (ALP, RUNX2/RUNX2, OSX/OSX, OCN/OCN, COL-I/COL-I, DSPP/DSP, and DMP-1/DMP-1) in IGF-1 treated SCAPs were significantly upregulated in Let-7c-low group. On the contrary, hsa-let-7c over-expression could downregulate the expression of these odonto/osteogenic markers. Moreover, western blot assay showed that the JNK and p38 MAPK signaling pathways were activated in Let-7c-low SCAPs but inhibited in Let-7c-over SCAPs. Together, the IGF-1/IGF-1R/hsa-let-7c axis can control the odonto/osteogenic differentiation of IGF-1-treated SCAPs via the regulation of JNK and p38 MAPK signaling pathways.

MicroRNA hsa‐let‐7b suppresses the odonto/osteogenic differentiation capacity of stem cells from apical papilla by targeting MMP1

“…Then, SCAPs were purified by the procedure of magnetic activated cell sorting (MACS) as previously described. 26 Cells were passaged by 0.25% trypsin at a 1:3 split ratio after the cell density came up to 80-85%. For the follow-up experiments, SCAPs within 3-5 passages were used.…”

“…The best parameter of multiplicity of infection (MOI) was 5 as previously reported. 26 SCAPs cultured at 60-70% confluence were transfected with the lentiviruses in 2 mL α-MEM supplemented with 10% FBS and 0.05 mg/mL polybrene (Santa Cruz Biotechnology, Santa Cruz, CA). Ten hours later, the infected cells were incubated in replaced conventional media for further use.…”

Comparative Secretome Analysis of Mesenchymal Stem Cells From Dental Apical Papilla and Bone Marrow During Early Odonto/Osteogenic Differentiation: Potential Role of Transforming Growth Factor-β2

“…(D,F) Western blotting analysis of expression of RUNX2, ALP, DSPP, and DMP1 of BMSCs after culturing in different medium at 7 days ( Figure 4D) and 14 days ( Figure 4F). (Ma et al, 2016;Wang et al, 2018;Zhang et al, 2019). To the best of our knowledge, there are no published systemic proteomic analyses that explore the extracellular environment of SCAPs during osteo/odontogenic differentiation.…”

Noncoding RNAs: new insights into the odontogenic differentiation of dental tissue-derived mesenchymal stem cells

“…In 2016, Ma et al [32] showed that the insulin-like growth factor-1 (IGF-1)/IGF-1R/hsa-let-7c axis exerts a key influence on the odontogenic differentiation of IGF-1-treated SCAPs as well as the MAPK signaling pathway. IGF-1 activity is mostly facilitated through IGF-1R and is therefore known as the IGF-1/IGF-1R axis.…”

“…Under these conditions, cells have been shown to subsequently express an osteoblast-associated gene profile, including alkaline phosphatase (ALP), collagen type 1 (COL-I), dentin matrix acid phosphoprotein 1 (DMP1), dentin sialophosphoprotein (DSPP), matrix extracellular phosphoglycoprotein (MEPE), osterix (OSX), osteocalcin (OCN), and osteopontin (OPN) [31]. Some of these genes regulate the expression of runtrelated transcription factor 2 (RUNX2), OSX, and COL-I at the early stage of odontoblast differentiation, while OCN participates in the later stage of differentiation [32]. The control of odontogenic differentiation of dental tissuederived MSCs shows great potential in the application of oral regenerative medicine and cytology treatment.…”