Oxytocin facilitates the proliferation, migration and osteogenic differentiation of human periodontal stem cells in vitro.

To explore the effect of oxytocin (OT) on the proliferation, migration, and osteogenic differentiation of periodontal ligament stem cells (PDLSCs) in vitro. PDLSCs were obtained by limiting dilution method. Immunofluorescence (IF), cell-counting kit-8 (CCK8), cell migration assay, Alizarin Red S staining, cetylpyridinium chloride (CPC) colorimetry, quantitative real-time polymerase chain reaction (qRT-PCR), and western blot analysis were used to examine the effect of OT on oxytocin receptor (OTR) expression, cell proliferation, migration and osteogenic differentiation of PDLSCs. Our study showed that PDLSCs expressed OTR. One hundred nanomolar OT exhibited the maximal effect on migration, while only 50 nM OT significantly promoted proliferation of PDLSCs, as well as mineralized nodule formation and calcium deposition (p < 0.05). Furthermore, 50 nM OT significantly up-regulated expression of osteogenesis-related genes-alkaline phosphatase (ALP), Collagen I (Col I), runt related transcription factor 2 (Runx 2), osteopontin (OPN) and osteocalcin (OCN)-at specific time points compared with osteogenic inductive medium (p < 0.05). In addition, western blot analysis demonstrated that 50 nM OT enhanced protein levels of ALP, Col I, and Runx 2 at day 7 and day 14 (p < 0.01), as well as activating the phosphorylation of extracellular-signal-regulated kinase (ERK) and protein kinase B (AKT) pathway; notably, 50 nM OT inhibited phosphorylation of the phosphatidylinositol 3-kinase (PI3K) pathway (p < 0.05). OT promoted proliferation, migration, and osteogenic differentiation of PDLSCs in vitro. Furthermore, the effect of OT on osteogenic differentiation was mediated through ERK and AKT pathway. Thus, OT may have potential for use in periodontal regeneration.

Ge B ，Liu H ，Liang Q ，Shang L ，Wang T ，Ge S ... -  《-》

Enamel matrix derivative enhances the proliferation and osteogenic differentiation of human periodontal ligament stem cells on the titanium implant surface.

Li G ，Hu J ，Chen H ，Chen L ，Zhang N ，Zhao L ，Wen N ，Yang Y ... -  《-》

PTH/SDF-1α cotherapy promotes proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells.

Stromal cell-derived factor-1α (SDF-1α) plays an important role in tissue regeneration in various tissues including the periodontium. A potential limitation for its use derives from its sensitivity to cleavage by dipeptidyl peptidase-IV (DPP-IV). Parathyroid hormone (PTH) reduces enzymatic activity of DPP-IV and is suggested to be a promising agent for periodontal tissue repair. The purpose of this study was to provide insight into how SDF-1α and intermittent PTH treatment might affect proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) in vitro. PDLSCs were isolated by the limiting dilution method. Surface markers were quantified by flow cytometry. Cell-counting kit-8 (CCK8), cell migration assay, alkaline phosphatase (ALP) activity assay, alizarin red staining and RT-PCR were used to determine viability, migration and osteogenic differentiation of PDLSCs. PDLSCs were positive for CD44, CD73, CD90, CD105, CD166 and STRO-1 and negative for CD14, CD34 and CD45. PTH/SDF-1α cotherapy significantly promoted cell proliferation, chemotactic capability, ALP activity and mineral deposition (P<.05). Gene expression level of bone sialoprotein (BSP), runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) were all up-regulated (P<.05). PTH/SDF-1α cotherapy promoted proliferation, migration and osteogenic differentiation of PDLSCs in vitro. Cotherapy seemed to have potential to promote periodontal tissue regeneration by facilitating chemotaxis of PDLSCs to the injured site, followed by promoting proliferation and osteogenic differentiation of these cells.

Du L ，Feng R ，Ge S 《-》

Potassium dihydrogen phosphate promotes the proliferation and differentiation of human periodontal ligament stem cells via nuclear factor kappa B pathway.

Periodontal ligament stem cells (PDLSCs) are vital for the regeneration of periodontal tissues. Potassium dihydrogen phosphate (KH2PO4) has recently been applied as a component of the mineralization inducing medium (MM), which can be used to induce osteogenic differentiation of dental stem cells. However, whether KH2PO4 has effects on PDLSCs has not been studied. PDLSCs were isolated by magnetic activated cell sorting and cultured. Alkaline phosphatase (ALP) activity and ALP protein expression of PDLSCs treated with different concentrations of KH2PO4 were examined to make sure the optimal concentration of KH2PO4 for the following experiments. The effects of KH2PO4 on the proliferation and differentiation of PDLSCs were investigated by flow cytometry, cell counting kit-8 assay, alizarin red staining, real-time RT-PCR, and Western blot. The involvement of nuclear factor kappa B (NF-κB) pathway in KH2PO4-treated PDLSCs was analyzed by Western blot and alizarin red staining. ALP activity assay and ALP protein expression examination revealed that 1.8 mmol/L KH2PO4 was the optimal concentration for the induction of hPDLSCs by KH2PO4. The proliferation and mineralization capacity of PDLSCs treated with KH2PO4 were enhanced as compared with the control group. PDLSCs treated with KH2PO4 showed an improved proliferation capacity in logarithmic growth phase at day 7. As PDLSCs were treated with KH2PO4, the expression of odonto/osteogenic markers (OCN/OCN, DSP/DSPP, OSX/OSX, RUNX2/RUNX2, and ALP/ALP) in cells were up-regulated at day 3 or 7. Moreover, the expression of IκBα in cytoplasm was down-regulated, along with an increased expression of p-P65 in cytoplasm and an up-regulated expression of P65 in nucleus. When treated with BMS345541 (the specific NF-κB inhibitor), the odonto/osteogenic differentiation of KH2PO4-treated PDLSCs was significantly attenuated. KH2PO4 can improve the proliferation and odonto/osteogenic differentiation capacity of PDLSCs via NF-κB pathway, and thus represents a potential target involved in the regeneration of periodontium for clinical treatments.

Xu Y ，Wang Y ，Pang X ，Li Z ，Wu J ，Zhou Z ，Xu T ，Gobin Beharee R ，Jin L ，Yu J ... -  《-》

Bone morphogenetic protein-9 induces PDLSCs osteogenic differentiation through the ERK and p38 signal pathways.

Ye G ，Li C ，Xiang X ，Chen C ，Zhang R ，Yang X ，Yu X ，Wang J ，Wang L ，Shi Q ，Weng Y ... -  《International Journal of Medical Sciences》