Enhanced Anti-inflammatory Capacity of the Conditioned Medium Derived from Periodontal Ligament Stem Cells Modified with an Iron-Based Nanodrug.

Cell-free therapy using conditioned medium (CM) from mesenchymal stem cells takes full advantage of the bioactive factors secreted by the cells while avoiding disadvantages such as immune rejection and tumor formation due to cell transplantation. In this study, human periodontal ligament stem cells (PDLSCs) are modified with the superparamagnetic iron oxide nanoparticle (SPION)-based nanodrug ferumoxytol (PDLSC-SPION). Compared with PDLSCs, PDLSC-SPION showed good cell viability and better osteogenic differentiation ability. Cell-free CM is collected and the anti-inflammatory capacity of PDLSC CM and PDLSC-SPION CM is assessed by treatment of lipopolysaccharide-stimulated macrophages and IL-17-stimulated human gingival fibroblasts. Both CMs inhibited the expression of proinflammatory cytokines in cells, and the therapeutic effect is more distinct for PDLSC-SPION CM than PDLSC CM, which may be due to their different proteomic compositions. Therefore, modification of PDLSCs with ferumoxytol enhances the anti-inflammatory capacity of its CM, making it more potentially useful for the treatment of inflammatory diseases such as periodontitis.

An altered oral microbiota induced by injections of superparamagnetic iron oxide nanoparticle-labeled periodontal ligament stem cells helps periodontal bone regeneration in rats.

Stem cell injection is good for periodontal regeneration due to the capacity of stem cells to differentiate toward osteogenic direction and to regulate the production of pro- and anti-inflammatory cytokines. However, injected cells are difficult to track in vivo. And there is microbiota in oral cavity, the dysbiosis of which leads to the damage and loss of periodontal tissue. Here, we demonstrated an enhanced periodontal repair was due to an altered oral microbiota. Periodontal defects were surgically prepared in rats, and periodontal ligament stem cells (PDLSCs) labeled by superparamagnetic iron oxide (SPIO) nanoparticles (PC-SPIO) were injected, with PDLSCs and saline treatments as controls. Detected by magnetic resonance imaging (MRI) and histological staining, PC-SPIO was major at limited areas in regenerated periodontal tissues. PC-SPIO-treated rats achieved better periodontal regeneration than the other two groups. Concurrently, the oral microbiota of PC-SPIO-treated rats was changed, presenting SPIO-Lac as a biomarker. SPIO-Lac assisted periodontal repair in vivo, inhibited the inflammation of macrophages induced by lipopolysaccharide (LPS) and antibacterial in vitro. Therefore, our study proved that SPIO-labeled cells can be tracked in periodontal defect and highlighted a potential positive role of an oral microbiota in periodontal regeneration, suggesting the possibility of periodontal repair promotion by manipulating oral microbiota.

A three-dimensional-printed SPION/PLGA scaffold for enhanced palate-bone regeneration and concurrent alteration of the oral microbiota in rats.

Scaffold implantation for the repair of oral bone defects involves an interplay between the scaffold biomaterial and the microenvironment. However, previous studies on this subject have only considered the effects of the immune system and largely ignored those of the oral microbiota. Accordingly, in the present study, we prepared composite scaffolds comprising a three-dimensional poly(l-lactide-co-glycolide) matrix with a superparamagnetic iron oxide nanoparticle (SPION) coating and used a rat model to evaluate their palate-bone-regenerating effects and their interaction with the oral microbiota. It was found that the SPION coated scaffold induced better bone regeneration than that achieved by the controls. Furthermore, it significantly decreased the operational taxonomic units (OTU) numbers as determined by 16 s rRNA gene sequencing, and also resulted in decreased Chao and ACE alpha diversity indexes compared with those of the controls. However, it had no effect on beta diversity. SPION coated scaffolds caused a shift in oral bacterial composition characterized by a decrease in the Clostridium spp. population, and the dominant flora being Proteobacteria. Furthermore, SPION coated scaffolds upregulated the concentration of serum iron, hepcidin, and P1NP. Thus, SPION coated scaffolds enhanced bone regeneration, and this effect was partly related to alteration of the oral microbiota by the antibacterial effects of SPION. Our findings provide a better understanding of the role of oral microbiota in oral bone regeneration and how SPION coated scaffolds can be used to enhance it.

3D magnetic nanocomposite scaffolds enhanced the osteogenic capacities of rat bone mesenchymal stem cells in vitro and in a rat calvarial bone defect model by promoting cell adhesion.

Magnetic scaffolds incorporated with iron oxide nanoparticles (IONPs) are biocompatible and present excellent osteogenic properties. However, the underlying mechanism is unclear. In this study, 3D-printed poly(lactic-co-glycolic acid) scaffolds were coated with IONPs using layer-by-layer assembly (Fe-scaffold) to prepare magnetic scaffolds. The effects of this modification on osteogenesis were investigated by comparison with untreated scaffolds (Uncoated-scaffold). The results showed that the proliferation of rat bone mesenchymal stem cells (rBMSCs) on the Fe-scaffold was enhanced compared with those on the Uncoated-scaffold (p < 0.05). The alkaline phosphatase activity and expression levels of osteogenic-related genes of cells on the Fe-scaffold were higher than those on the Uncoated-scaffold (p < 0.05). Fe-scaffold was found to promote the cell adhesion compared with Uncoated-scaffold, including increasing the adhered cell number, promoting cell spreading and upregulating the expression levels of adhesion-related genes integrin α1 and ß1 and their downstream signaling molecules FAK and ERK1/2 (p < 0.05). Moreover, the amount of new bone formed in rat calvarial defects at 8 weeks decreased in the order: Fe-scaffold > Uncoated-scaffold > Blank-control (samples whose defects were left empty) (p < 0.05). Therefore, 3D magnetic nanocomposite scaffolds enhanced the osteogenic capacities of rBMSCs in vitro and in a rat calvarial bone defect model by promoting cell adhesion. The mechanisms were attributed to the alteration in its hydrophilicity, surface roughness, and chemical composition.