Simvastatin induces the osteogenic differentiation of human periodontal ligament stem cells

Abstract

Periodontal ligament stem cells (PDLSCs) are considered as potential mesenchymal stem cell sources for future clinical applications in periodontal regeneration therapy. Simvastation, widely used for lowering serum cholesterol, is known to have a bone stimulatory effect. However, it is not clear whether simvastation affects the differentiation of PDLSCs. This study examined the effects of simvastatin on human PDLSCs in vitro and in vivo. Using the limiting dilution technique, human PDLSCs were isolated and expanded. PDLSCs were cultured with simvastatin (0.01-10 μM), and the proliferation was measured. The osteogenic differentiation was characterized by alkaline phosphatase (ALP) activity and Alizarin Red-S staining for calcium deposition. The gene expression levels of osteogenic markers were evaluated by RT-PCR. In addition, PDLSCs were transplanted into nude mice with ceramic bovine bone powders as carriers to observe the capacity of mineralized tissue formation in vivo. Simvastatin at concentrations <1 μM did not suppress the proliferation of PDLSCs. After the administration of 0.1 μM simvastatin, the expression of ALP, bone sialoprotein, and bone morphogenetic protein-2 genes were significantly upregulated, and the ALP activity and mineralized nodule formation were significantly higher in the simvastatin-treated cells than the control cells. In addition, the in vivo transplantation results showed that simvastatin treatment promoted the degree of mineralized tissue formation. Collectively, simvastatin has positive effects on osteogenic differentiation of human PDLSCs in vitro and in vivo. This suggests that simvastatin might be a useful osteogenic induction agent for periodontal bone regeneration.

Simvastatin Induces In Vitro Mineralization Effects of Primary Human Odontoblast-Like Cells

“…Significantly decreased cell viability, increased phosphate turnover and significant ( p < 0.01) increase of the mineralization capability of the odontoblast-like cells point to a linkage between SV and the ability of biomineralization of odontoblast-like cells especially at a later stage of culture in the current study (day 21) ( Figure 4 ) [ 30 , 34 , 35 , 37 ]. Although cytotoxic effects of SV are recognizable in a time and dose-dependent manner upon human osteoblasts, odontoblast-like cells and likewise other cell types [ 23 , 24 , 46 , 47 ] the application of SV induced a 2 times elevated median value of the ALP conversion in the group with 2 µM SV compared to the control group ( Figure 3 ). Additionally, the median value of the mineralization capability of the group with the same concentration was approximately 3 times higher compared with the control group ( Figure 4 ).…”

Statins in nonsurgical and surgical periodontal therapy. A systematic review and meta‐analysis of preclinical in vivo trials

“…Local application instead of systemic administration is also supported by the following facts: (i) 10%‐25% of patients taking statins systemically report muscle symptoms, are exposed to an increased risk for myopathy and hepatotoxicity and also present a slightly higher risk for developing diabetes mellitus; (ii) these side effects are dose‐dependent; (iii) neither herein nor in clinical studies have any remarkable adverse reactions been reported after local statin application, except for some local postoperative inflammation; and (iv) no systemic effects of local application have been detected (e.g., no alterations in the nearby submandibular lymph nodes, liver enzyme levels, numbers of white blood cells and immune cells and body weight). Nevertheless, it must be mentioned that local application of high statin concentrations may also exert an antiproliferative and even an apoptotic effect on periodontal cell types (i.e., gingival fibroblasts, periodontal ligament stem cells); therefore, there is space for optimization of dose and delivery systems.…”

Evaluation of Bone Regeneration of Simvastatin Loaded Chitosan Nanofiber Membranes in Rodent Calvarial Defects

“…Simvastatin has the ability to upregulate BMP-2 mRNA expression in human mesenchymal stem cells [36]. In addition, osteogenic differentiation of human periodontal ligament stem cells cultured with simvastatin was enhanced and characterized by elevated expression levels of osteogenic markers such as alkaline phosphatase, bone sialoprotein, and BMP-2 [37]. In fact, simvastatin has the ability to antagonize tumor necrosis factor-alpha (TNF-α), an inflammatory cytokine that inhibits osteogenesis, and subsequently upregulate the expression of BMP-2 in osteoblasts [38].…”