The Synergistic Effect of Electrical Stimulation and Dermal Fibroblast Cells-Laden 3D Conductive Hydrogel for Full-Thickness Wound Healing.

Clinically, most patients with poor wound healing suffer from generalized skin damage, usually accompanied by other complications, so developing therapeutic strategies for difficult wound healing has remained extremely challenging until now. Current studies have indicated that electrical stimulation (ES) to cutaneous lesions enhances skin regeneration by activating intracellular signaling cascades and secreting skin regeneration-related cytokine. In this study, we designed different concentrations of graphene in gelatin-methacrylate (GelMa) to form the conductive composite commonly used in wound healing because of its efficiency compared to other conductive thermo-elastic materials. The results demonstrated the successful addition of graphene to GelMa while retaining the original physicochemical properties of the GelMa bioink. In addition, the incorporation of graphene increased the interactions between these two biomaterials, leading to an increase in mechanical properties, improvement in the swelling ratio, and the regulation of degradation characteristics of the biocomposite scaffolds. Moreover, the scaffolds exhibited excellent electrical conductivity, increasing proliferation and wound healing-related growth factor secretion from human dermal fibroblasts. Overall, the HDF-laden 3D electroconductive GelMa/graphene-based hydrogels developed in this study are ideal biomaterials for skin regeneration applications in the future.

Mechanical assessment and odontogenic behavior of a 3D-printed mesoporous calcium silicate/calcium sulfate/poly-ε-caprolactone composite scaffold.

BACKGROUND/PURPOSE: Tissue engineering in dentistry has fundamentally changed the way endodontists assess treatment options. Our previous study found that quercetin-contained mesoporous calcium silicate/calcium sulfate (MSCSQ) could induce hard tissue defect region regeneration. This study focused on whether the MSCSQ scaffold could also be effective in regulating odontogenesis and dentin regeneration. METHODS: In this study, we fabricated MSCSQ composite scaffolds using the 3D printing technique. The characteristics of the MSCSQ scaffold were examined by scanning electron microscope (SEM), and mechanical properties were also assessed. In addition, we evaluated the cell viability, cell proliferation, odontogenic-related protein expression, and mineralization behavior of human dental pulp stem cells (hDPSCs) cultured on different scaffolds. RESULTS: We found the precipitation of spherical-apatite on the scaffold surface rapidly in short periods. The in-vitro results for cell behavior revealed that hDPSCs with an MSCSQ scaffold were significantly higher in cell viability as followed time points. In addition, the specific makers of odontogenesis, such as DSPP and DMP-1 proteins, were induced obviously after culturing the hDPSCs on the MSCSQ scaffold. CONCLUSION: Our results demonstrated that MSCSQ scaffolds could enhance physicochemical and biological behaviors compared to mesoporous calcium silicate/calcium sulfate (MSCS) scaffolds. In addition, MSCSQ scaffolds also enhanced odontogenic and immuno-suppressive properties compared to MSCS scaffolds. These results indicated that MSCSQ scaffolds could be considered a potential bioscaffold for clinical applications and dentin regeneration.

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.

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.

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.

Incorporation of Calcium Sulfate Dihydrate into a Mesoporous Calcium Silicate/Poly-ε-Caprolactone Scaffold to Regulate the Release of Bone Morphogenetic Protein-2 and Accelerate Bone Regeneration.

Tissue engineering and scaffolds play an important role in tissue regeneration by supporting cell adhesion, proliferation, and differentiation. The design of a scaffold is critical in determining its feasibility, and it is critical to note that each tissue is unique in terms of its morphology and composition. However, calcium-silicate-based scaffolds are undegradable, which severely limits their application in bone regeneration. In this study, we developed a biodegradable mesoporous calcium silicate (MS)/calcium sulfate (CS)/poly-ε-caprolactone (PCL) composite and fabricated a composite scaffold with 3D printing technologies. In addition, we were able to load bone morphogenetic protein-2 (BMP-2) into MS powder via a one-step immersion procedure. The results demonstrated that the MS/CS scaffold gradually degraded within 3 months. More importantly, the scaffold exhibited a gradual release of BMP-2 throughout the test period. The adhesion and proliferation of human dental pulp stem cells on the MS/CS/BMP-2 (MS/CS/B) scaffold were significantly greater than that on the MS/CS scaffold. It was also found that cells cultured on the MS/CS/B scaffold had significantly higher levels of alkaline phosphatase activity and angiogenic-related protein expression. The MS/CS/B scaffold promoted the growth of new blood vessels and bone regeneration within 4 weeks of implantation in rabbits with induced critical-sized femoral defects. Therefore, it is hypothesized that the 3D-printed MS/CS/B scaffold can act both as a conventional BMP-2 delivery system and as an ideal osteoinductive biomaterial for bone regeneration.

Mineral Trioxide Aggregate Mixed with 5-Aminolevulinic Acid for the Photodynamic Antimicrobial Strategy in Hard Tissue Regeneration.

Aminolevulinic acid (ALA) based photodynamic antimicrobial strategy can provide good antimicrobial effects and be used for medical applications. The aim of this study was to apply this strategy to Mineral Trioxide Aggregate (MTA), which is commonly used as a filling material for root endings and by doing so, to increase the bactericidal capability of MTA, as well as to investigate its characterization, cytocompatibility, and odontogenic differentiation potential. MTA is known to be a derivative of calcium silicate (CS). In this study, MTA specimens with or without ALA and light treatment were prepared. Diametral tensile strength values (DTS), setting durations, X-ray diffraction (XRD) spectra, apatite-mineralization, and antimicrobial abilities of the MTA, were also analyzed. Human dental pulp cells (hDPCs) can proliferate into the newly formed matrix and differentiate into odontoblasts to reinforce and strengthen the root. Levels of hDPCs proliferation and its odontogenic capabilities when cultured on MTA with ALA and light treatment, and the percentages of cells existing in the various cell cycle stages, were further evaluated in this study. The results indicated that MTA added ALA with light treatment had greater antibacterial ability and cytocompatibility, compared to MTA alone. A higher percentage S phase of the cells cultured on MTA added ALA with light treatment was observed. Furthermore, hDPCs cultured on MTA added ALA with light treatment had the highest expression levels of the odontoblastic differentiation markers. ALA has great antimicrobial efficiency and is a potential material for future medical applications. ALA-based photodynamic antibacterial strategy applied in the MTA has great antibacterial ability, cytocompatibility, and odontoblastic differentiation potential, and can facilitate the development of root canal treatment.

Mineral Trioxide Aggregate with Mussel-inspired Surface Nanolayers for Stimulating Odontogenic Differentiation of Dental Pulp Cells.

INTRODUCTION: This research was intended to evaluate the feasibility of mineral trioxide aggregate (MTA) powder coated with polydopamine (PDA) in dental and bone tissue regeneration by investigating the hydration, physicochemical properties, and biological performance of hydrated cements. METHODS: The MTA powder was well suspended in a dopamine solution buffered at a pH of 8.5 using tris(hydroxymethyl)aminomethane buffer and vigorously stirred for 12 hours at room temperature. The PDA-coated MTA powder was mixed with water and hydrated at 37°C with 100% relative humidity for 1 day. The setting time, mechanical strength, phase composition, surface morphology, and in vitro bioactivity of the cements as well as the proliferation and odontogenic differentiation of human dental pulp cells cultured on the cements were evaluated. RESULTS: The setting of the MTA cements was significantly shortened without jeopardizing the mechanical properties with PDA incorporated into the cements. In addition, our results proved that PDA-coated MTA up-regulation of odontogenic-related protein of hDPCs. PDA-coated MTA induced the odontogenic differentiation of cells as indicated by an alkaline phosphate activity test and an odontogenic-related protein analysis. CONCLUSIONS: These results indicate that dopamine is an effective coating material to promote long-term human dental pulp cell culture and odontogenic differentiation on PDA-MTA substrates. This will be an important direction for future studies focused on developing new biomaterials for dental applications.

Surface Modification of Calcium Silicate via Mussel-Inspired Polydopamine and Effective Adsorption of Extracellular Matrix to Promote Osteogenesis Differentiation for Bone Tissue Engineering.

Calcium silicate-based cement has garnered huge interest in recent years, due to its versatility and potential in mass fabrication of a variety of bioceramics. For this study, the main objective was to fabricate functionalized calcium silicate (CS) powder integrated with a simple bio-inspired surface modification using polydopamine (PDA), to regulate cellular behaviors such as cellular adhesion, and subsequently cell differentiation and proliferation. For this study, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) techniques were used to analyze the chemical compositions and observe the surface characteristics of our PDA coated CS cements. Such modifications were found to enhance Wharton Jelly's mesenchymal stem cells (WJMSC) in various ways. Firstly, PDA-coated CS cements were found to significantly enhance cell adhesion with higher expressions of cell adhesion markers, such as focal adhesion kinase and integrins. This was further supported by morphology analysis of the cells. This enhanced cell adhesion, in turn, led to significantly higher secretion of extracellular matrix (ECM) proteins, such as collagen I and fibronectin, which directly promoted cell attachments and proliferation. In our osteogenesis assays, it was found that secretion and expression of osteogenesis related genes and proteins were significantly higher and were dependent on the PDA content. Therefore, these results demonstrated that such simple bio-inspired modification techniques of synthetic degradable CS cements can be applied as a future modification, to modify and convert inert surfaces of synthetic bone grafts to enhance and modulate the cell behaviors of WJMSCs. This in turn can be used as a potential alternative for further bioengineering research.

Laser Sintered Magnesium-Calcium Silicate/Poly-ε-Caprolactone Scaffold for Bone Tissue Engineering.

In this study, we manufacture and analyze bioactive magnesium-calcium silicate/poly-ε-caprolactone (Mg-CS/PCL) 3D scaffolds for bone tissue engineering. Mg-CS powder was incorporated into PCL, and we fabricated the 3D scaffolds using laser sintering technology. These scaffolds had high porosity and interconnected-design macropores and structures. As compared to pure PCL scaffolds without an Mg-CS powder, the hydrophilic properties and degradation rate are also improved. For scaffolds with more than 20% Mg-CS content, the specimens become completely covered by a dense bone-like apatite layer after soaking in simulated body fluid for 1 day. In vitro analyses were directed using human mesenchymal stem cells (hMSCs) on all scaffolds that were shown to be biocompatible and supported cell adhesion and proliferation. Increased focal adhesion kinase and promoted cell adhesion behavior were observed after an increase in Mg-CS content. In addition, the results indicate that the Mg-CS quantity in the composite is higher than 10%, and the quantity of cells and osteogenesis-related protein of hMSCs is stimulated by the Si ions released from the Mg-CS/PCL scaffolds when compared to PCL scaffolds. Our results proved that 3D Mg-CS/PCL scaffolds with such a specific ionic release and good degradability possessed the ability to promote osteogenetic differentiation of hMSCs, indicating that they might be promising biomaterials with potential for next-generation bone tissue engineering scaffolds.

Fabrication and characterization of polycaprolactone and tricalcium phosphate composites for tissue engineering applications.

BACKGROUND/PURPOSE: ß-Tricalcium phosphate (ß-TCP) is an osteoconductive material which has been used for clinical purposes for several years, as is polycaprolactone (PCL), which has already been approved for a number of medical and drug delivery devices. In this study we have incorporated various concentrations of ß-TCP into PCL with the aim of developing an injectable, mechanically strong, and biodegradable material which can be used for medical purposes without organic solvents. MATERIALS AND METHODS: This study assesses the physical and chemical properties of this material, evaluates the in vitro bioactivity of the PCL/ß-TCP composites, and analyzes cell proliferation and osteogenic differentiation when using human bone marrow mesenchymal stem cells (hBMSCs). RESULTS: The results show that weight losses of approximately 5.3%, 12.1%, 18.6%, and 25.2%, were observed for the TCP0, TCP10, TCP30, and TCP50 composites after immersion in simulated body fluid for 12 weeks, respectively, indicating significant differences (P < 0.05). In addition, PCL/ß-TCP composites tend to have lower contact angles (47 ± 1.5° and 58 ± 1.7° for TCP50 and TCP30, respectively) than pure PCL (85 ± 1.3°), which are generally more hydrophilic. After 7 days, a significant (22% and 34%, respectively) increase (P < 0.05) in alkaline phosphatase level was measured for TCP30 and TCP50 in comparison with the pure PCL. CONCLUSION: PCL/TCP is biocompatible with hBMSCs. It not only promotes proliferation of hBMSCs but also helps to differentiate reparative hard tissue. We suggest 50% (weight) PCL-containing ß-TCP biocomposites as the best choice for hard tissue repair applications.

The Characteristics of Mineral Trioxide Aggregate/Polycaprolactone 3-dimensional Scaffold with Osteogenesis Properties for Tissue Regeneration.

INTRODUCTION: The aim of this study was to investigate whether the mineral trioxide aggregate/polycaprolactone (MTA/PCL) hybrid 3-dimensional (3D) scaffold supplies a suitable microenvironment for the osteogenic differentiation of human dental pulp cells (hDPCs) and to further consider the effect of the MTA/PCL composite on the biological performance of hybrid scaffolds. METHODS: MTA was suspended in absolute alcohol and dropped slowly into PCL that was generated with the printable MTA-matrix. Then, the MTA/PCL composite was prepared into highly uniform scaffolds with controlled macropore sizes and structure using a 3D printing technique. Mechanical properties and the apatite precipitation of the scaffolds were evaluated as well as the cell response to the scaffolds by culturing hDPCs. RESULTS: The results showed that the MTA/PCL 3D scaffold had uniform, 450-µm, high-porosity (70%) macropores and a compressive strength of 4.5 MPa. In addition, the MTA/PCL scaffold could effectively promote the adhesion, proliferation, and differentiation of hDPCs. CONCLUSIONS: The 3D-printed MTA/PCL scaffolds not only exhibited excellent physical and chemical properties but also enhanced osteogenesis differentiation. All of the results support the premise that this MTA/PCL porous scaffold would be a useful biomaterial for application in bone tissue engineering.

Calcium Silicate/Chitosan-Coated Electrospun Poly (Lactic Acid) Fibers for Bone Tissue Engineering.

Electrospinning technology allows fabrication of nano- or microfibrous fibers with inorganic and organic matrix and it is widely applied in bone tissue engineering as it allows precise control over the shapes and structures of the fibers. Natural bone has an ordered composition of organic fibers with dispersion of inorganic apatite among them. In this study, poly (lactic acid) (PLA) mats were fabricated with electrospinning and coated with chitosan (CH)/calcium silicate (CS) mixer. The microstructure, chemical component, and contact angle of CS/CH-PLA composites were analyzed by scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. In vitro, various CS/CH-coated PLA mats increased the formation of hydroxyapatite on the specimens' surface when soaked in cell cultured medium. During culture, several biological characteristics of the human mesenchymal stem cells (hMSCs) cultured on CS/CH-PLA groups were promoted as compared to those on pure PLA mat. Increased secretion levels of Collagen I and fibronectin were observed in calcium silicate-powder content. Furthermore, with comparison to PLA mats without CS/CH, CS10 and CS15 mats markedly enhanced the proliferation of hMSCs and their osteogenesis properties, which was characterized by osteogenic-related gene expression. These results clearly demonstrated that the biodegradable and electroactive CS/CH-PLA composite mats are an ideal and suitable candidate for bone tissue engineering.

Preparation of the fast setting and degrading Ca-Si-Mg cement with both odontogenesis and angiogenesis differentiation of human periodontal ligament cells.

Develop a fast setting and controllable degrading magnesium-calcium silicate cement (Mg-CS) by sol-gel, and establish a mechanism using Mg ions to stimulate human periodontal ligament cells (hPDLs) are two purposes of this study. We have used the diametral tensile strength measurement to obtain the mechanical strength and stability of Mg-CS cement; in addition, the cement degradation properties is realized by measuring the releasing amount of Si and Mg ions in the simulated body fluid. The other cell characteristics of hPDLs, such as proliferation, differentiation and mineralization were examined while hPDLs were cultured on specimen surfaces. This study found out the degradation rate of Mg-CS cements depends on the Mg content in CS. Regarding in vitro bioactivity; the CS cements were covered with abundant clusters of apatite spherulites after immersion of 24h, while less apatite spherulites were formatted on the Mg-rich cement surfaces. In addition, the authors also explored the effects of Mg ions on the odontogenesis and angiogenesis differentiation of hPDLs in comparison with CS cement. The proliferation, alkaline phosphatase, odontogenesis-related genes (DSPP and DMP-1), and angiogenesis-related protein (vWF and ang-1) secretion of hPDLs were significantly stimulated when the Mg content of the specimen was increased. The results in this study suggest that Mg-CS materials with this modified composition could stimulate hPDLs behavior and can be good bioceramics for bone substitutes and hard tissue regeneration applications as they stimulate odontogenesis/angiogenesis.

The Ionic Products from Mineral Trioxide Aggregate-induced Odontogenic Differentiation of Dental Pulp Cells via Activation of the Wnt/ß-catenin Signaling Pathway.

INTRODUCTION: Mineral trioxide aggregate (MTA) has been successfully used in clinical applications in endodontics. However, little is known about the involvement of Wnt/ß-catenin signaling in human dental pulp cells (hDPC) differentiation with the interaction of MTA in hard tissue regeneration, especially in odontogenesis. Therefore, the aim of this study was to explore odontogenic/osteogenic gene expression and the protein secretion of hDPCs cultured with consecutive concentrations of MTA extracts and carefully examine the particular molecular mechanism that occurs during this process. METHODS: MTA extracts were prepared by immersing MTA powders into Dulbecco modified Eagle medium at a concentration of 200 mg/mL. hDPCs were cultured with various concentrations of MTA extracts, and the resulting changes in the cells, such as proliferation and odontogenic differentiation, were measured. RESULTS: The results indicate that hDPC proliferation increases remarkably in a time-dependent manner in most treatment groups, except the highest concentration group (200 mg/mL). The Wnt/ß-catenin signaling pathway-related genes and proteins are significantly raised when hDPCs are cultured in a wide concentration range of MTA extracts compared with a control, except for the highest concentration group (100 mg/mL), on days 3 and 7 (P < .05). CONCLUSIONS: These variations indicate that Wnt/ß-catenin signaling is involved in MTA extract-induced odontogenic differentiation of hDPCs.

Physical characteristics, antimicrobial and odontogenesis potentials of calcium silicate cement containing hinokitiol.

Hinokitiol is a natural material and it has antibacterial and anti-inflammatory effects. The purpose of this study was to evaluate the material characterization, cell viability, antibacterial and anti-inflammatory abilities of the hinokitiol-modified calcium silicate (CS) cement as a root end filling material. The setting times, diametral tensile strength (DTS) values and XRD patterns of CS cements with 0-10mM hinokitiol were examined. Then, the antibacterial effect and the expression levels of cyclooxygenase 2 (COX-2) and interleukin-1 (IL-1) of the hinokitiol-modified CS cements were evaluated. Furthermore, the cytocompatibility, the expression levels of the markers of odontoblastic differentiation, mineralized nodule formation and calcium deposition of human dental pulp cells (hDPCs) cultured on hinokitiol-modified CS cements were determined. The hinokitiol-modified CS cements had better antibacterial and anti-inflammatory abilities and cytocompatibility than non-modified CS cements. Otherwise, the hinokitiol-modified CS cements had suitable setting times and better odontoblastic potential of hDPCs. Previous report pointed out that the root-end filling materials may induce inflammatory cytokines reaction. In our study, hinokitiol-modified CS cements not only inhibited the expression level of inflammatory cytokines, but also had better cytocompatibility, antimicrobial properties and active ability of odontoblastic differentiation of hDPCs. Therefore, the hinokitiol-modified CS cement may be a potential root end filling material for clinic.

Human Dental Pulp Cells Responses to Apatite Precipitation from Dicalcium Silicates.

Unraveling the mechanisms behind the processes of cell attachment and the enhanced proliferation that occurs as a response to the presence of calcium silicate-based materials needs to be better understood so as to expand the applications of silicate-based materials. Ions in the environment may influence apatite precipitation and affect silicate ion release from silicate-based materials. Thus, the involvement of apatite precipitate in the regulation of cell behavior of human dental pulp cells (hDPCs) is also investigated in the present study, along with an investigation of the specific role of cell morphology and osteocalcin protein expression cultured on calcium silicate (CS) with different Dulbecco's modified Eagle's medium (DMEM). The microstructure and component of CS cement immersion in DMEM and P-free DMEM are analyzed. In addition, when hDPCs are cultured on CS with two DMEMs, we evaluate fibronectin (FN) and collagen type I (COL) secretion during the cell attachment stage. The facilitation of cell adhesion on CS has been confirmed and observed both by scanning with an electron microscope and using immunofluorescence imaging. The results indicate that CS is completely covered by an apatite layer with tiny spherical shapes on the surface in the DMEM, but not in the P-free DMEM. Compared to the P-free DMEM, the lower Ca ion in the DMEM may be attributed to the formation of the apatite on the surfaces of specimens as a result of consumption of the Ca ion from the DMEM. Similarly, the lower Si ion in the CS-soaked DMEM is attributed to the shielding effect of the apatite layer. The P-free DMEM group releases more Si ion increased COL and FN secretion, which promotes cell attachment more effectively than DMEM. This study provides new and important clues regarding the major effects of Si-induced cell behavior as well as the precipitated apatite-inhibited hDPC behavior on these materials.

Antibacterial and Odontogenesis Efficacy of Mineral Trioxide Aggregate Combined with CO2 Laser Treatment.

INTRODUCTION: Mineral trioxide aggregate (MTA) has been successfully used in clinical applications in endodontics. Studies show that the antibacterial effects of CO2 laser irradiation are highly efficient when bacteria are embedded in biofilm because of a photothermal mechanism. The aim of this study was to confirm the effects of CO2 laser irradiation on MTA with regard to both material characterization and cell viability. METHODS: MTA was irradiated with a dental CO2 laser using directly mounted fiber optics in the wound healing mode with a spot area of 0.25 cm(2) and then stored in an incubator at 100% relative humidity and 37°C for 1 day to set. The human dental pulp cells cultured on MTA were analyzed along with their proliferation and odontogenic differentiation behaviors. RESULTS: The results indicate that the setting time of MTA after irradiation by the CO2 laser was significantly reduced to 118 minutes rather than the usual 143 minutes. The maximum diametral tensile strength and x-ray diffraction patterns were similar to those obtained without CO2 laser irradiation. However, the CO2 laser irradiation increased the amount of Ca and Si ions released from the MTA and regulated cell behavior. CO2 laser-irradiated MTA promoted odontogenic differentiation of hDPCs, with the increased formation of mineralized nodules on the substrate's surface. It also up-regulated the protein expression of multiple markers of odontogenic and the expression of dentin sialophosphoprotein protein. CONCLUSIONS: The current study provides new and important data about the effects of CO2 laser irradiation on MTA with regard to the decreased setting time and increased ion release. Taking cell functions into account, the Si concentration released from MTA with laser irradiation may be lower than a critical value, and this information could lead to the development of new regenerative therapies for dentin and periodontal tissue.