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.

Coupling Biocompatible Au Nanoclusters and Cellulose Nanofibrils to Prepare the Antibacterial Nanocomposite Films.

Cellulose nanofibrils (CNF) is considered as an inexhaustible precursor to produce antibacterial materials, such as antibacterial hydrogel, antibacterial paper, and antibacterial film. However, the poor antimicrobial property of neat CNF required it should be coupled with an antibacterial ingredient. Herein, biocompatible Au nanoclusters (AuNCs) were synthesized and added into the CNF dispersion to prepare a novel antibacterial film (AuNCs@CNF film). The effects of addition of AuNCs with different amount on the morphology and physicochemical properties of AuNCs@CNF films were characterized using atomic force microscopy (AFM), scanning electron microscopy (SEM), X-ray diffraction (XRD), FTIR (Fourier-transform infrared), light transmittance spectra, and thermogravimetric analysis (TGA). The results showed that AuNCs did not affect the nano-structural features of the CNF film and its basic structures, but could greatly increase the hydrophilicity, the flexibility and the thermal stability of CNF film, which might improve its application in antimicrobial wound-healing dressing. The prepared AuNCs@CNF films demonstrated high antibacterial properties toward Escherichia coli (E. coli) and Streptococcus mutans (S. mutans) both in vitro and in vivo, which can prohibit their growths and promote the healing of bacteria-infected wound, respectively. Thus, the prepared AuNCs@CNF film with great antibacterial properties could be applicable in biomedical field.

Iron oxide nanoparticle-calcium phosphate cement enhanced the osteogenic activities of stem cells through WNT/ß-catenin signaling.

Calcium phosphate cement (CPC), functionalized with iron oxide nanoparticles (IONP), is of great promise to promote osteoinduction and new bone formation. In this work, the IONP powder was added into the CPC powder to fabricate CPC + IONP scaffolds and the effects of the novel composite on bone matrix formation and osteogenesis of human dental pulp stem cells (hDPSCs) were explored. A series of CPC + IONP magnetic scaffolds with different IONP contents (1%, 3% and 6%) were fabricated using 5% chitosan solution as the cement liquid. Western blotting and RT-PCR were used to analyze the signaling pathway. The IONP incorporation substantially enhanced the performance of CPC + IONP, with increases in both mechanical strength and cellular activities. The IONP addition greatly promoted the osteogenesis of hDPSCs, elevating the ALP activity, the expression of osteogenic marker genes and bone matrix formation with 1.5-2-fold increases. The 3% IONP incorporation showed the most enhancement among all groups. Activation of the extracellular signal-related kinases WNT/ß-catenin in DPSCs was observed, and this activation was attenuated by the WNT inhibitor DKK1. The results indicated that the osteogenic behavior of hDPSCs was likely driven by CPC + IONP via the WNT signaling pathway. In conclusion, incorporate IONP into CPC scaffold remarkably enhanced the spreading, osteogenic differentiation and bone mineral synthesis of stem cell. Therefore, this method had great potential for bone tissue engineering. The novel CPC + IONP composite scaffolds with stem cells are promising to provide an innovative strategy to enhance bone regenerative therapies.

Biocompatibility and osteogenic activity of guided bone regeneration membrane based on chitosan-coated magnesium alloy.

Ideally, a guided bone regeneration membrane (GBRM) should possess high strength, as for titanium membranes, along with excellent biocompatibility and osteoconductivity, as for natural absorbable collagen membranes. Besides titanium, magnesium (Mg) is another metal widely used in the biomedical field, which also exhibits biodegradability. In this study, a composite chitosan­magnesium (CS-Mg) membrane was fabricated by dip-coating Mg alloy into chitosan solution. In vitro and in vivo tests were performed to investigate whether this membrane could be used as biodegradable GBRM, and the test results were compared with those obtained for a commercial GBRM (Heal-All). The microstructure was analyzed by scanning electron microscopy-electron dispersive spectroscopy. The degradation behavior was investigated by immersing the membranes into Dulbecco's modified Eagle medium (DMEM). The in vitro biocompatibility was evaluated by cell adhesion, cytotoxicity and alkaline phosphatase (ALP) assays using MG63 cells. The cytotoxicity and ALP assays were performed with diluted extracts of Mg, CS-Mg and Heal-All. The results show that CS-Mg has a suitable degradation rate, as well as similar cell adhesion and cytocompatibility to Heal-All. However, the 10% CS-Mg extracts exhibited higher ALP activity at 3 and 5 days (p < 0.05) compared with the medium control and the Heal-All extracts, but no differences with 10% Mg extracts (p > 0.05). Rabbit calvarial defects were used for testing the osteogenic activity in vivo. Three groups of samples were examined: CS-Mg, Heal-All, and a blank control. Higher amounts of new bone were formed for the CS-Mg and Heal-All groups (p < 0.05) compared with the blank control, whereas no significant differences between the CS-Mg and Heal-All groups were observed (p > 0.1). In conclusion, the CS-Mg membrane shows great potential for application as a biodegradable metallic GBRM with excellent osteogenic activity.

Enhanced osteoinduction of electrospun scaffolds with assemblies of hematite nanoparticles as a bioactive interface.

PURPOSE: Electrospun scaffolds have been studied extensively for their potential use in bone tissue engineering. However, their hydrophobicity and relatively low matrix stiffness constrain their osteoinduction capacities. In the present study, we studied polymer electrospun scaffolds coated with hydrophilic hematite nanoparticles (αFeNPs) constructed using layer-by-layer (LbL) assembly to construct a bioactive interface between the scaffolds and cells, to improve the osteoinduction capacities of the scaffolds. MATERIALS AND METHODS: The morphology of the αFeNPs was assessed. Surface properties of the scaffolds were tested by X-ray photoelectron spectroscopy (XPS), surface water contact angle, and in vitro protein adsorption test. The stiffness of the coating was tested using an atomic force microscope (AFM). In vitro cell assays were performed using rat adipose-derived stem cells (ADSCs). RESULTS: Morphology characterizations showed that αFeNPs assembled on the surface of the scaffold, where the nano assemblies improved hydrophilicity and increased surface roughness, with increased surface stiffness. Enhanced initial ADSC cell spread was found in the nano assembled groups. Significant enhancements in osteogenic differentiation, represented by enhanced alkaline phosphatase (ALP) activities, elevated expression of osteogenic marker genes, and increased mineral synthesis by the seeded ADSCs, were detected. The influencing factors were attributed to the better hydrophilicity, rougher surface topography, and harder interface stiffness. In addition, the presence of nanoparticles was believed to provide better cell adhesion sites. CONCLUSION: The results suggested that the construction of a bioactive interface by LbL assembly using αFeNPs on traditional scaffolds should be a promising method for bone tissue engineering.