The aim of this study was to assess the efficacy of a self-assembling peptide hydrogel as a scaffold for bone regeneration. We used a neutral and injectable self-assembling peptide hydrogel, SPG-178-Gel. Bone defects (5 mm in diameter) in rat calvarial bones were filled with a mixture of alpha-modified Eagle's medium and peptide hydrogel. Three weeks after surgery, soft X-ray and microcomputed tomography (micro-CT) images of the gel-treated bones showed new bone formations in the periphery and in central areas of the defects. Next, we evaluated the three-dimensional osteogenic induction of dental pulp stem cells (DPSCs), a type of mesenchymal stem cell, in SPG-178-Gel. We first confirmed that the osteogenic differentiation of DPSCs was significantly promoted by osteogenic induction medium containing recombinant human bone morphogenetic protein-4 (rhBMP-4) in a two-dimensional cell culture. Then, we verified DPSC proliferation and osteogenic differentiation in a three-dimensional cell culture using SPG-178-Gel. The gene expression levels of osteopontin, osteocalcin, and collagen type I were significantly increased when DPSCs were cultured in SPG-178-Gel with the osteogenic induction medium. Micro-CT observations showed the formation of widespread calcium deposition. In conclusion, SPG-178-Gel was adequately effective as a scaffold and can be a suitable tool for bone formation in vivo and in vitro. These findings suggest that the self-assembling peptide hydrogel, SPG-178-Gel, could be a promising tool for bone tissue engineering.
To develop a root canal filling materialwith high antimicrobial activity, we prepared guttapercha supplemented with the cationic surfactant cetylpyridinium chloride (CPC). Thermoplastic gutta-percha was supplemented with 0.05%, 0.2%, or 0.8% CPC. The gutta-percha containing CPC was tightly packed at the bottom of a 24-well plate. Its antimicrobial activity against eight representative endodontic pathogens-including gram-positive and gram-negative bacteria and fungi-was evaluated by adding 0.5 mL of liquid samples containing pathogens to the wells. After 24 h of cultivation under appropriate conditions, microbial growth was analyzed by counting colony-forming units (CFU). Gutta-percha alone (without CPC) partially inhibited microbial growth, probably through the antimicrobial effect of some of its components, such as zinc oxide. Addition of CPC dose-dependently increased the antimicrobial efficacy of gutta-percha. Addition of 0.05%, 0.2%, and 0.8% CPC reduced the viable microbial number to below the lower limit of detection (20 CFU/mL) for all tested pathogens except Pseudomonas aeruginosa, which was detected in 0.8% CPC-containing guttapercha, although the viable number significantly decreased. Gutta-percha with CPC might be useful for preventing microbial infections during root canal therapy. (J Oral Sci 58, 277-282, 2016)
In order to develop an self-cured acrylic resin having an antibacterial property, three types of commercially available inorganic antibacterial agents were added, at 1% each, to UNIFAST III to evaluate the antibacterial property. The antibacterial test evaluated the amount of Streptococcus mutans attached to UNIFAST III, the residual viable count of Streptococcus mutans cultured on UNIFAST III. And the color tone changes evaluated immediate and temporal color tone changes of UNIFAST III caused by the addition of antibacterial agents. As a result, compared to UNIFAST III without any added inorganic antibacterial agent, a significant decrease was observed in the attachment amount and the residual viable count of Streptococcus mutans. In addition, when adding NOVARON from among the added inorganic antibacterial agents, UNIFAST III exhibited little change in the temporal color tone. Thus, these results suggest that the addition of antibacterial agents to resins is effective first step toward developing self-cured acrylic resins having an antibacterial property.
Residual monomers in resin-based biomaterials cause cytotoxicity. We previously showed that methyl methacrylate (MMA) induced mRNA expression of the glutathione S-transferase alpha 1 gene (Gsta1) located downstream of the cis-acting anti-oxidant responsive element (ARE). Herein, we tested the hypothesis that MMA activated the Gsta1 promoter through the ARE. HepG2 cells were transfected with a luciferase reporter vector containing the ARE and the Gsta1 promoter (-990 to +46 bp) and cultured for 12 hrs with MMA (initial concentration, 10 mM). Analysis of the expressed luciferase activity indicated that MMA activated the promoter 2.6-fold. MMA (from 1 to 30 mM) dose-dependently increased the promoter activity, which reached a plateau between 6 and 12 hrs. In HepG2 cells transfected with a reporter vector containing 2 AREs and a TATA-like promoter, 10 mM MMA increased the reporter expression 2.8-fold. These results suggest that MMA increases Gsta1 transcription through ARE-mediated promoter activation.
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