ABSTRACT. Periodontal ligament stem cells (PDLSCs) are tissue-specific mesenchymal stem cells (MSCs), having an important role in regenerative therapy for teeth loss. Interleukin-7 (IL-7) is a key cytokine produced by stromal cells including MSCs, and exhibits specific roles for B and T cell development and osteoblasts differentiation of multiple myeloma. However, the effect of IL-7 on osteogenic differentiation of PDLSCs remains unclear. Therefore, in the present study we determined whether IL-7 affects the proliferation and osteogenic differentiation of PDLSCs in vitro and explored the associated signaling pathways for IL-7-mediated cell differentiation. The results demonstrated that the isolated human PDLSCs possessed MSCs features, highly expressing CD90, CD44, CD105, CD29 and CD73, and almost did not expressed CD34, CD45, CD11b, CD14 and CD117. IL-7 could not significantly affect the proliferation of PDLSCs, but it decreased their osteogenic differentiation and inhibited alkaline phosphatase (ALP) activity. The results of quantitative reverse transcriptionpolymerase chain reaction (qRT-PCR) and Western blotting exhibited that the expression levels of Runx-2, SP7 and osteocalcin (OCN) were significantly reduced by IL-7. Further studies indicated that IL-7 did not significantly change JNK, ERK1/2 and p38 protein production, but markedly suppressed their phosphorylation levels. These data suggest that IL-7 inhibits the osteogenic differentiation of PDLSCs probably via inactivation of mitogen-activated protein kinase (MAPK) signaling pathway.
Ethnopharmacological relevance: Gualou Xiebai Banxia decoction (GLXBBX) is a well-known traditional Chinese herbal formula that was first discussed in the Synopsis of the Golden Chamber by Zhang Zhongjing in the Eastern Han Dynasty. In traditional Chinese medicine, GLXBBX is commonly prescribed to treat cardiovascular diseases, such as coronary heart disease and atherosclerosis. Objective: The present study aimed to examine GLXBBX’s preventative capacity and elucidate the potential molecular mechanism of Poloxamer 407 (P407)-induced hyperlipidemia in rats. Materials and Methods: Both the control and model groups received pure water, and the test group also received a GLXBBX decoction. For each administration, 3 mL of the solution was administered orally. To establish hyperlipidemia, a solution mixed with 0.25 g/kg P407 dissolved in 0.9% normal saline was injected slowly into the abdominal cavity. At the end of the study, the rats’ plasma lipid levels were calculated using an automatic biochemical analyzer to evaluate the preventative capability of the GLXBBX decoction, and the serum and liver of the rats were collected. Results: The GLXBBX decoction significantly improved P407-induced hyperlipidemia, including increased plasma triglycerides, aspartate aminotransferase elevation, and lipid accumulation. Moreover, GLXBBX decoction treatment increased lipoprotein lipase (LPL) activity and mRNA expression of LPL. Furthermore, GLXBBX significantly suppressed the mRNA expression of stearoyl-CoA desaturase (SCD1). Conclusion: GLXBBX significantly improved P407-induced hyperlipidemia, which may have been related to enhanced LPL activity, increased LPL mRNA expression, and decreased mRNA expression of SCD1.
The present study aims to investigate the effect of recombinant human bone
morphogenetic protein-2 (rhBMP-2) on the osteogenesis of periodontal ligament (PDL)
cells. The expression vector of rhBMP-2 (pcDNA3.1-rhBMP-2) was established. PDL cells
were obtained through the enzymatic digestion and tissue explant methods and verified
by immunohistochemistry. Cells were classified into experimental (cells transfected
with pcDNA3.1/rhBMP-2-EGFP), blank (cells with no transfection) and control
group (cells transfected with empty plasmid). rhBMP-2 expression was assessed via
Western blotting analysis. The mineralization ability, alkaline phosphatase (ALP)
activity and level of related osteogenic biomarkers were detected to evaluate the
osteogenic characteristics of PDL cells. The rhBMP-2 expression vector
(pcDNA3.1-rhBMP-2) was successfully established. Primary PDL cells displayed a star
or long, spindle shape. The cultured cells were long, spindle-shaped, had a plump
cell body and homogeneous cytoplasm and the ellipse nucleus contained two or three
nucleoli. Cells displayed a radial, sheaf-like or eddy-like arrangement after
adherence growth. Immunohistochemical staining confirmed that cells originated from
mesenchymal opposed to epithelium. The experimental group exhibited an enhanced
mineralization ability, higher ALP activity and increased expression of rhBMP-2 and
osteogenic biomarkers (Runx2, collagen type I and osteocalcin) than the blank and
control group. The present study demonstrated that rhBMP-2 transfection enhances the
osteogenesis of PDL cells and provides a possibility for the application of rhBMP-2
expression products in dental disease treatment.
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