Cytokines regulate stemness of mesenchymal stem cells via miR‐628‐5p during periodontal regeneration

Abstract

Background Cytokines play key roles in stimulating periodontal regeneration; however, their exact mechanisms of action remain unclear. Mesenchymal stem cells (MSCs) are multipotent cells that have self‐renewal abilities and can differentiate into periodontal tissues such as bone, cementum, and periodontal ligaments following transplantation, like periodontal progenitor cells. Here, we used MSCs to identify the regulatory genes induced by periodontal regenerative cytokines. Methods Human MSCs (hMSCs) were cultured under conditions of periodontal regenerative cytokine stimulation or silencing of undifferentiated hMSC transcription factors. To characterize the changes associated with periodontal regenerative cytokine‐regulated microRNAs (miRNAs), miRNA, and mRNA expression was evaluated using miRNA arrays and quantitative real‐time polymerase chain reaction, respectively. One of the identified miRNAs, miR‐628‐5p, was then overexpressed or suppressed in hMSCs during osteogenesis; the effect of these changes on osteogenesis was investigated. Results Cytokine‐stimulated MSCs showed characteristic miRNA profiles and mRNA levels of undifferentiated hMSC transcription factors ETV1, SOX11, and GATA6. Next, we silenced these transcription factors in MSCs and examined the miRNA profiles. The levels of miR‐628‐5p were decreased upon all cytokine treatments and were increased upon silencing of ETV1, SOX11, and GATA6. Overexpression of miR‐628‐5p suppressed osteogenesis; however, its inhibition enhanced OPN, ALP, OC, BMP2, and RUNX2 mRNA levels, and bone matrix mineralization, but not OSX mRNA or ALP activity. Conclusions miR‐628‐5p negatively regulates MSC stemness during periodontal regeneration. Periodontal regenerative cytokines act as miR‐628‐5p suppressors to support periodontal regeneration. Thus, selection of effective cytokines for different MSCs, based on miRNA profiling, is important for advancing regenerative therapies.

Periodontal tissue regeneration with cementogenesis after application of brain‐derived neurotrophic factor in 3‐wall inflamed intra‐bony defect

“…These cytokines characteristically enhance cementoblast-related or periodontal ligament cell-related genes and microRNAs. 60,61 Therefore, the function of BDNF and the other cytokines can be distinguished in the periodontal regenerative process and effect, and these cytokines can creatively be applied based on each morphology of bone defects.…”

“…Moreover, BDNF, as well as some other cytokines applied for periodontal regeneration therapy, regulates the cementogenesis of undifferentiating multiple progenitor cells, for example, mesenchymal stem cells. These cytokines characteristically enhance cementoblast‐related or periodontal ligament cell‐related genes and microRNAs 60,61 . Therefore, the function of BDNF and the other cytokines can be distinguished in the periodontal regenerative process and effect, and these cytokines can creatively be applied based on each morphology of bone defects.…”

Advances Focusing on the Application of Decellularized Extracellular Matrix in Periodontal Regeneration

“…For example, deoxyprostaglandin nanoparticles were found to be anti-inflammatory and improve regeneration by Jiang et al [ 7 ]. Cytokines such as BMP-2, FGF, VEGF, and TGF-β were normally added in periodontal regeneration with positive results, and the combination of cytokines and other bioactive extracts with dECM materials achieved a greater outcome in both bone tissue engineering and periodontium regeneration [ 57 , 58 , 59 , 61 , 62 ].…”

Regulation of osteogenesis in bone marrow‐derived mesenchymal stem cells via histone deacetylase 1 and 2 co‐cultured with human gingival fibroblasts and periodontal ligament cells

“…17 Several genes have been reported as regulatory factors in MSC osteogenesis, including basic helix-loop-helix (bHLH) transcription factors, Twist-related protein 1 (TWIST1), 18 differentiated embryonic chondrocyte expressed gene 1 (DEC1), 19 and DEC2, 20 and MSC undifferentiating transcription factors. 21,22 Additionally, microR-NAs, such as miR-383, 14 miR-628-5p, 14,23 miR-29, 24 miR-204, 25 and miR-335 26 are implicated in MSC osteogenesis.…”