Substitutions of strontium in bioactive calcium silicate bone cements stimulate osteogenic differentiation in human mesenchymal stem cells.

Calcium silicate cements have been considered as alternative bone substitutes owing to its extraordinary bioactivity and osteogenicity. Unfortunately, the major disadvantage of the cements was the slow degradation rate which may limit the efficiency of bone regeneration. In this study, we proposed a facile method to synthesize degradable calcium silicate cements by incorporating strontium into the cements through solid-state sintering. The effects of Sr incorporation on physicochemical and biological properties of the cements were evaluated. Although, our findings revealed that the incorporation of strontium retarded the hardening reaction of the cements, the setting time of different cements (11-19 min) were in the acceptable range for clinical use. The presence of Sr in the CS cements would hampered the precipitation of calcium phosphate products on the surface after immersion in SBF, however, a layer of precipitated calcium phosphate products can be formed on the surface of the Sr-CS cement within 1 day immersion in SBF. More importantly, the degradation rate of the cements increased with increasing content of strontium, consequentially raised the levels of released strontium and silicon ions. The elevated dissolving products may contribute to the enhancement of the cytocompatibility, alkaline phosphatase activity, osteocalcin secretion, and mineralization of human Wharton's jelly mesenchymal stem cells. Together, it is concluded that the strontium-incorporated calcium silicate cement might be a promising bone substitute that could accelerate the regeneration of irregularly shaped bone defects.

Trigeminocardiac reflex during non-surgical root canal treatment of teeth with irreversible pulpitis.

BACKGROUND/PURPOSE: Trigeminocardiac reflex (TCR) is a unique clinical incident of acute change in hemodynamic balance, which may lead to hypotension, bradycardia, and even clinical crisis. Up to date, no study so far considers the impact of non-surgical root canal treatment (NSRCT) of irreversible pulpitis teeth under either local infiltration or block anesthesia on hemodynamic change possibly related to TCR. METHODS: This study enrolled 111 patients with 138 irreversible pulpitis teeth that were treated by two sessions of NSRCT. The first session involved mainly the removal of vital pulp tissue with the direct stimulation of the dental branches of the trigeminal nerve, and the second session included the root canal enlargement and debridement with minimal disturbance to the dental branches of the trigeminal nerve. Vital signs mainly the blood pressure were recorded during both NSRCT sessions. RESULTS: The incidences of NSRCT patients with MABP decrease ≧10%, ≧15%, or ≧20% were all significantly higher in the first NSRCT session than in the second NSRCT session (all the P-values < 0.001). In the first NSRCT session, the incidence of patients with MABP decrease ≧10% was significantly associated with tooth type. For both upper and lower teeth, the patients with premolars treated by NSRCR had significantly higher incidences of MABP decrease ≧10% than those with either anterior or molar teeth treated by NSRCR (all the P-values < 0.05). CONCLUSION: We conclude that vital pulp extirpation may lead to a substantial drop in patient's blood pressure possibly related to TCR.

Tensile force on human macrophage cells promotes osteoclastogenesis through receptor activator of nuclear factor κB ligand induction.

Little is known about the effects of tensile forces on osteoclastogenesis by human monocytes in the absence of mechanosensitive cells, including osteoblasts and fibroblasts. In this study we consider the effects of tensile force on osteoclastogenesis in human monocytes. The cells were treated with receptor activator of nuclear factor κB ligand (RANKL) to promote osteoclastogenesis. Then,expression and secretion of cathepsin K were examined. RANKL and the formation of osteoclasts during the osteoclast differentiation process under continual tensile stress were evaluated by Western blot. It was also found that -100 kPa or lower induces RANKL-enhanced tartrate-resistant acid phosphatase activity in a dose-dependent manner. Furthermore, an increased tensile force raises the expression and secretion of cathepsin K elevated by RANKL, and is concurrent with the increase of TNF-receptor-associated factor 6 induction and nuclear factor κB activation. Overall, the current report demonstrates that tensile force reinforces RANKL-induced osteoclastogenesis by retarding osteoclast differentiation. The tensile force is able to modify every cell through dose-dependent in vitro RANKL-mediated osteoclastogenesis, affecting the fusion of preosteoclasts and function of osteoclasts. However, tensile force increased TNF-receptor-associated factor 6 expression. These results are in vitro findings and were obtained under a condition of tensile force. The current results help us to better understand the cellular roles of human macrophage populations in osteoclastogenesis as well as in alveolar bone remodeling when there is tensile stress.

The synergistic effects of Chinese herb and injectable calcium silicate/ß-tricalcium phosphate composite on an osteogenic accelerator in vitro.

This study investigates the physicochemical and biological effects of traditional Chinese medicines on the ß-tricalcium phosphate (ß-TCP)/calcium silicate (CS) composites of bone cells using human dental pulp cell. CS is an osteoconductive and bioactive material. For this research we have combined ß-TCP and CS and check its effectiveness, a series of ß-TCP/CS composites with different ratios of Xu Duan (XD) were prepared to make new bioactive and biodegradable biocomposites for bone repair. XD has been used in Traditional Chinese Medicine for hundreds of years as an antiosteoporosis, tonic and antiaging agent for the therapy of low back pain, traumatic hematoma, threatened abortion and bone fractures. Formation of bone-like apatite, the diametral tensile strength, and weight loss of composites were considered before and after immersion in simulated body fluid (SBF). In addition, we also examined the effects of XD released from ß-TCP/CS composites and in vitro human dental pulp cell (hDPCs) and studied its behavior. The results show the XD-contained paste did not give any demixing when the weight ratio of XD increased to 5-10 % due to the filter-pressing effect during extrusion through the syringe. After immersion in SBF, the microstructure image showed a dense bone-like apatite layer covered on the ß-TCP/CS/XD composites. In vitro cell experiments shows that the XD-rich composites promote human dental pulp cells (hDPCs) proliferation and differentiation. However, when the XD quantity in the composite is more than 5 %, the amount of cells and osteogenesis protein of hDPCs were stimulated by XD released from ß-TCP/CS composites. The combination of XD in degradation of ß-TCP and osteogenesis of CS gives strong reason to believe that these calcium-based composite cements may prove to be promising bone repair materials.

Activation of focal adhesion kinase induces extracellular signal-regulated kinase-mediated osteogenesis in tensile force-subjected periodontal ligament fibroblasts but not in osteoblasts.

The exact mechanism by which focal adhesion kinase (FAK) translates mechanical signals into osteogenesis differentiation in force-subjected cells has not been elucidated. The responses to different forces differ according to the origin of cells and the type of stress applied. Therefore, the recruitment of osteoclast and osteoblast progenitor cells, and the balanced activation of these cells around and within the periodontal ligament (PDL) are essential for alveolar bone remodeling. Cells within the PDL and MG63 cells were subjected to tensile forces of -100 kPa for different periods of time. At various times during the tensile force application, they were processed for the purpose of analyzing cell viability, cell cycle, and osteogenic protein. The effect of small interfering RNA transfection targeting FAK was also evaluated. Tensile force enhanced a rapid increase in the phosphorylation of FAK and up-regulated osteogenic protein expression in PDL cells, but not in MG63 cells. Transfecting PDL cells with FAK antisense oligonucleotide diminished alkaline phosphatase and osteocalcin secretion. These findings suggest that tensile force activates FAK pathways in PDL cells, which down-regulate immune cytokine and up-regulate osteogenic protein.

Low-level diode laser therapy reduces lipopolysaccharide (LPS)-induced bone cell inflammation.

UNLABELLED: In this study, the aim is to investigate the cytologic effects of inflammatory bone cells after in vitro low-level laser therapy (LLLT). A human osteosarcoma cell line (MG63) was cultured, infected with lipopolysaccharide (LPS) and exposed to low-level laser treatment at 5 or 10 J/cm(2) using a 920 nm diode laser. MG63 cell attachment was observed under a microscope, and cell viability was quantified by mitochondrial colorimetric assay (MTT). LPS-treated MG63 cells were irradiated with LLLT, and the inflammatory markers iNOS, TNF-α and IL-1, were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. The data were collected and analyzed by one-way analysis of variance (ANOVA); p < 0.05 indicated a statistically significant difference. Low-level laser treatment on MG63 cells increased their ability to attach and survive. After irradiation, the expression levels of iNOS, TNF-α and IL-1 in LPS-infected MG63 cells decreased over time (p < 0.05). CONCLUSIONS: low-level diode laser treatment increased the MG63 cell proliferative ability and decreased the expression of inflammatory mediators in MG63 cells.

An in vitro comparison of the frictional forces between archwires and self-ligating brackets of passive and active types.

The aim of this study was to compare the static and kinetic frictional forces generated by various contemporary designs of self-ligating brackets (SLBs) and different wire alloys. In total, six different brackets (four passive type SLB, one active SLB, and one conventional bracket) were investigated using stainless steel, nickel-titanium, and titanium-molybdenum alloy archwires of several sizes. The friction forces were measured by sliding on a bracket-wire combination system in an EZ instron testing machine. A scanning electron microscope (SEM) was used to examine the wear effects of the wall surfaces of bracket slots. Energy-dispersive spectroscopy (EDS) was used to identify the elemental compositions of the bracket surfaces. The data were collected and statistically analysed using analysis of variance. The results of static and kinetic frictional forces were lower in passive type SLBs (P < 0.05), except in the Smart Clip bracket. The wire materials or wire dimensions in the present study showed similar friction forces with no statistical differences (P > 0.05). The wearing effects were not obviously found in bracket slots under SEM observation. Only conventional brackets and mini-Clippy SLB revealed nickel ions via EDS analysis. This study shows that passive SLBs are associated with lower static or kinetic friction forces than those of active SLBs or conventional brackets. Wear on the bracket slots was not observed in the present study.

Synergistic Effect of Static Magnetic Fields and 3D-Printed Iron-Oxide-Nanoparticle-Containing Calcium Silicate/Poly-ε-Caprolactone Scaffolds for Bone Tissue Engineering.

In scaffold-regulated bone regeneration, most three-dimensional (3D)-printed scaffolds do not provide physical stimulation to stem cells. In this study, a magnetic scaffold was fabricated using fused deposition modeling with calcium silicate (CS), iron oxide nanoparticles (Fe3O4), and poly-ε-caprolactone (PCL) as the matrix for internal magnetic sources. A static magnetic field was used as an external magnetic source. It was observed that 5% Fe3O4 provided a favorable combination of compressive strength (9.6 ± 0.9 MPa) and degradation rate (21.6 ± 1.9% for four weeks). Furthermore, the Fe3O4-containing scaffold increased in vitro bioactivity and Wharton's jelly mesenchymal stem cells' (WJMSCs) adhesion. Moreover, it was shown that the Fe3O4-containing scaffold enhanced WJMSCs' proliferation, alkaline phosphatase activity, and the osteogenic-related proteins of the scaffold. Under the synergistic effect of the static magnetic field, the CS scaffold containing Fe3O4 can not only enhance cell activity but also stimulate the simultaneous secretion of collagen I and osteocalcin. Overall, our results demonstrated that Fe3O4-containing CS/PCL scaffolds could be fabricated three dimensionally and combined with a static magnetic field to affect cell behaviors, potentially increasing the likelihood of clinical applications for bone tissue engineering.

Comparison of friction force between corroded and noncorroded titanium nitride plating of metal brackets.

INTRODUCTION: Titanium nitride (TiN) plating is a method to prevent metal corrosion and can increase the surface smoothness. The purpose of this study was to evaluate the friction forces between the orthodontic bracket, with or without TiN plating, and stainless steel wire after it was corroded in fluoride-containing solution. METHODS: In total, 540 metal brackets were divided into a control group and a TiN-coated experimental group. The electrochemical corrosion was performed in artificial saliva with 1.23% acidulated phosphate fluoride (APF) as the electrolytes. Static and kinetic friction were measured by an EZ-test machine (Shimadazu, Tokyo, Japan) with a crosshead speed of 10 mm per minute over a 5-mm stretch of stainless steel archwire. The data were analyzed by using unpaired t test and analysis of variance (ANOVA). RESULTS: Both the control and TiN-coated groups' corrosion potential was higher with 1.23% APF solution than with artificial solution (P <0.05). In brackets without corrosion, both the static and kinetic friction force between the control and TiN-coated brackets groups showed a statistically significant difference (P <0.05). In brackets with corrosion, the control group showed no statistical difference on kinetic or static friction. The TiN-coated brackets showed a statistical difference (P <0.05) on kinetic and static friction in different solutions. CONCLUSION: TiN-coated metal brackets, with corrosion or without corrosion, cannot reduce the frictional force.

Bidirectional Differentiation of Human-Derived Stem Cells Induced by Biomimetic Calcium Silicate-Reinforced Gelatin Methacrylate Bioink for Odontogenic Regeneration.

Tooth loss or damage is a common problem affecting millions of people worldwide, and it results in significant impacts on one's quality of life. Dental regeneration with the support of stem cell-containing scaffolds has emerged as an alternative treatment strategy for such cases. With this concept in mind, we developed various concentrations of calcium silicate (CS) in a gelatin methacryloyl (GelMa) matrix and fabricated human dental pulp stem cells (hDPSCs)-laden scaffolds via the use of a bioprinting technology in order to determine their feasibility in promoting odontogenesis. The X-ray diffraction and Fourier transform-infrared spectroscopy showed that the incorporation of CS increased the number of covalent bonds in the GelMa hydrogels. In addition, rheological analyses were conducted for the different concentrations of hydrogels to evaluate their sol-gel transition temperature. It was shown that incorporation of CS improved the printability and printing quality of the scaffolds. The printed CS-containing scaffolds were able to release silicate (Si) ions, which subsequently significantly enhanced the activation of signaling-related markers such as ERK and significantly improved the expression of odontogenic-related markers such as alkaline phosphatase (ALP), dentin matrix protein-1 (DMP-1), and osteocalcin (OC). The calcium deposition assays were also significantly enhanced in the CS-containing scaffold. Our results demonstrated that CS/GelMa scaffolds were not only enhanced in terms of their physicochemical behaviors but the odontogenesis of the hDPSCs was also promoted as compared to GelMa scaffolds. These results demonstrated that CS/GelMa scaffolds can serve as cell-laden materials for future clinical applications and use in dentin regeneration.

Synergies of Human Umbilical Vein Endothelial Cell-Laden Calcium Silicate-Activated Gelatin Methacrylate for Accelerating 3D Human Dental Pulp Stem Cell Differentiation for Endodontic Regeneration.

According to the Centers for Disease Control and Prevention, tooth caries is a common problem affecting 9 out of every 10 adults worldwide. Dentin regeneration has since become one of the pressing issues in dentistry with tissue engineering emerging as a potential solution for enhancing dentin regeneration. In this study, we fabricated cell blocks with human dental pulp stem cells (hDPSCs)-laden alginate/fish gelatin hydrogels (Alg/FGel) at the center of the cell block and human umbilical vascular endothelial cells (HUVEC)-laden Si ion-infused fish gelatin methacrylate (FGelMa) at the periphery of the cell block. 1H NMR and FTIR results showed the successful fabrication of Alg/FGel and FGelMa. In addition, Si ions in the FGelMa were noted to be bonded via covalent bonds and the increased number of covalent bonds led to an increase in mechanical properties and improved degradation of FGelMa. The Si-containing FGelMa was able to release Si ions, which subsequently significantly not only enhanced the expressions of angiogenic-related protein, but also secreted some cytokines to regulate odontogenesis. Further immunofluorescence results indicated that the cell blocks allowed interactions between the HUVEC and hDPSCs, and taken together, were able to enhance odontogenic-related markers' expression, such as alkaline phosphatase (ALP), dentin matrix phosphoprotein-1 (DMP-1), and osteocalcin (OC). Subsequent Alizarin Red S stain confirmed the benefits of our cell block and demonstrated that such a novel combination and modification of biomaterials can serve as a platform for future clinical applications and use in dentin regeneration.

The synergistic effects of Xu Duan combined Sr-contained calcium silicate/poly-ε-caprolactone scaffolds for the promotion of osteogenesis marker expression and the induction of bone regeneration in osteoporosis.

Osteoporosis and its related problems such as fractures are gradually becoming common due to an aging population. Current methods to treat osteoporosis include medical and surgical options such as bone implants. Recent developments in 3D printing and materials science technologies has allowed us to fabricate individualized scaffolds with desired properties. In this study, we mixed Xu Duan into strontium­calcium silicate powder at 5% (XD5) and 10% (XD10) and fabricated 3D scaffolds with polycaprolactone. All scaffolds were assessed for its physical, mechanical, and biological properties to evaluated for its feasibility for bone tissue engineering in the osteoporosis model. Our results showed that such a scaffold could be fabricated using extrusion-based printing techniques and that addition of XD did not alter original structural properties of the SrCS. Furthermore, the XD5 and XD10 scaffolds were found to be non-toxic to cells and cells cultured on the scaffolds had significantly higher proliferation and secreted increased osteogenic-related proteins in in vitro studies as compared to the XD0 groups. Remarkably, the XD10 scaffolds could be used as substitutes for the critical-sized bone defect (7.0 mm diameter and 8.0 mm depth) in the osteoporotic rabbit model. The XD10 scaffolds can enhance bone ingrowth and accelerate new bone regeneration even in complex osteoporotic pathological environments. These results showed that such a Chinese medicine-contained scaffold had potential in osteoporosis bone tissue regeneration and could be considered as a promising tool for future clinical used applications.

The Development of Light-Curable Calcium-Silicate-Containing Composites Used in Odontogenic Regeneration.

Pulp regeneration is one of the most successful areas in the field of tissue regeneration, despite its current limitations. The biocompatibility of endodontic biomaterials is essential in securing the oral microenvironment and supporting pulp tissue regeneration. Therefore, the objective of this study was to investigate the new light-curable calcium silicate (CS)-containing polyethylene glycol diacrylate (PEGDA) biocomposites' regulation of human dental pulp stem cells (hDPSCs) in odontogenic-related regeneration. The CS-containing PEGDA (0 to 30 wt%) biocomposites are applied to endodontics materials to promote their mechanical, bioactive, and biological properties. Firstly, X-ray diffraction and Fourier-transform infrared spectroscopy showed that the incorporation of CS increased the number of covalent bonds in the PEGDA. The diameter tension strength of the CS-containing PEGDA composite was significantly higher than that of normal PEGDA, and a different microstructure was detected on the surface. Samples were analyzed for their surface characteristics and Ca/Si ion-release profiles after soaking in simulated body fluid for different periods of time. The CS30 group presented better hDPSC adhesion and proliferation in comparison with CS0. Higher values of odontogenic-related biomarkers were found in hDPSCs on CS30. Altogether, these results prove the potential of light-curable CS-containing PEGDA composites as part of a 'point-of-care' strategy for application in odontogenesis-related regeneration.

Variations in surface characteristics and corrosion behaviour of metal brackets and wires in different electrolyte solutions.

The aim of this study was to assess the surface characteristics and to compare the corrosion potential of metal brackets and wires in environments containing different media. Four brands of metal brackets and two types of orthodontic wires [stainless steel and nickel-titanium (NiTi)] were investigated. An electrochemical assay was used to compare the corrosion potential (V) of the brackets and wires in different electrolyte media at 37°C. The test media were acidulated sodium fluoride (NaF) and pH 4 and pH 6 artificial saliva solutions. The data were analysed using analysis of variance with a predetermined significance level of α = 0.05. Scanning electron microscopy (SEM) was used to observe surface defects and corrosion. The results of the potentiodynamic curve showed that most brands of metal brackets were easily corroded in the NaF and pH 4 environments, while the NiTi and stainless steel wires were easily corroded in the pH 4 artificial saliva. SEM observations showed that defects or pitting corrosion occurred on the surfaces of the brackets and wires in all tested media.

Caffeic Acid-coated Nanolayer on Mineral Trioxide Aggregate Potentiates the Host Immune Responses, Angiogenesis, and Odontogenesis.

INTRODUCTION: The aim of this study was to investigate whether mineral trioxide aggregate (MTA) can be modified with caffeic acid (CA) to form caffeic acid/mineral trioxide aggregate (CAMTA) cement and to evaluate its physicochemical and biological properties as well as its capability in immune suppression and angiogenesis. METHODS: MTA was immersed in trishydroxymethyl aminomethane buffer with CA to allow coating onto MTA powders. X-ray diffractometry and tensile stress-strain tests were conducted to assess for physical characteristics of CAMTA and to evaluate for successful modification of MTA. Then, the CAMTA cement was immersed in simulated body fluid to evaluate its hydroxyapatite formation capabilities and Si release profiles. In addition, RAW 264.7 cells and human dental pulp stem cells were used to evaluate CAMTA's immunosuppressive capabilities and cell responses, respectively. hDPSCs were also used to assess CAMTA's angiogenic capabilities. RESULTS: The X-ray diffractometry results showed that CA can be successfully coated onto MTA without disrupting or losing MTA's original structural properties, thus allowing us to retain the initial advantages of MTA. CAMTA was shown to have higher mechanical properties compared with MTA and had rougher pitted surfaces, which were hypothesized to lead to enhanced adhesion, proliferation, and secretion of angiogenic- and odontogenic-related proteins. In addition, it was found that CAMTA was able to enhance hydroxyapatite formation and immunosuppressive capabilities compared with MTA. CONCLUSIONS: CAMTA cements were found to have improved physicochemical and biological characteristics compared with their counterpart. In addition, CAMTA cements had enhanced odontogenic, angiogenic, and immunosuppressive properties compared with MTA. All of the results of this study proved that CAMTA cements could be a biomaterial for future clinical applications and tissue engineering use.

Orthodontic adhesives induce human gingival fibroblast toxicity and inflammation.

OBJECTIVE: To test the null hypothesis that the resin base and the resin hybrid glass ionomer base adhesives do not cause inflammation after contacting primary human gingival fibroblasts in vitro. MATERIALS AND METHODS: The resin base and resin hybrid glass ionomer base adhesives were used to treat human gingival fibroblasts to evaluate the survival rate using MTT colorimetric assay to detect the level of cyclooxygenase-2 (COX-2) mRNA by reverse transcription polymerase chain reaction (RT-PCR) technique and COX-2 protein expression using Western blot analysis. The results were analyzed using one-way analysis of variance (ANOVA). Tests of differences of the treatments were analyzed using the Tukey test and a value of P < .05 was considered statistically significant. RESULTS: The paste and primer of the resin base adhesive and the liquid of glass ionomer adhesive showed decreasing survival rates after 24 hours of treatment (P < .05). All orthodontic adhesives induced COX-2 protein expression in human gingival fibroblasts. The exposure of quiescent human gingival fibroblasts to adhesives resulted in the induction of COX-2 mRNA expression. The investigations of the time-dependent COX-2 mRNA expression in adhesive-treated human gingival fibroblasts revealed different patterns. CONCLUSIONS: The hypothesis is rejected. For orthodontic patients with gingival inflammation, except for those with oral hygiene problems, the activation of COX-2 expression by orthodontic adhesive may be one of the potential mechanisms.