Optimization by mixture design of chitosan/multi-phase calcium phosphate/BMP-2 biomimetic scaffolds for bone tissue engineering.

The modulation of cell behavior during culture is one of the most important aspects of bone tissue engineering because of the necessity for a complex mechanical and biochemical environment. This study aimed to improve the physicochemical properties of chitosan/multi-phase calcium phosphate (MCaP) scaffolds using an optimized mixture design experiment and evaluate the effect of biofunctionalization of the obtained scaffolds with the bone morphogenetic protein BMP-2 on stem cell behavior. The present study evaluated the compressive strength, elastic modulus, porosity, pore diameter, and degradation in simulated body fluids and integrated these responses using desirability. The properties of the scaffolds with the best desirability (18.4% of MCaP) were: compressive strength of 23 kPa, elastic modulus of 430 kPa, pore diameter of 163 µm, porosity of 92%, and degradation of 20% after 21 days. Proliferation and differentiation experiments were conducted using dental pulp stem cells after grafting BMP-2 onto scaffolds via the carbodiimide route. These experiments showed that MCaP promoted cell proliferation and increased alkaline phosphatase activity, whereas BMP-2 enhanced cell differentiation. This study demonstrates that optimizing the composition of a mixture of chitosan and MCaP improves the physicochemical and biological properties of scaffolds, indicating that this solution is viable for application in bone tissue engineering.

Microstructural, microchemical, and mechanical changes associated with the clinical reuse of two nickel-titanium endodontic instruments.

BACKGROUND: Nickel-titanium (NiTi) instruments have represented a great technological development that enabled endodontists conforming irregular-shaped root canals. Notwithstanding, the repeated use of these instruments may lead to the fracture without any prior visible warning signs. This study aimed to evaluate how multiple clinical instrumentation/sterilization cycles of two NiTi mechanized instruments can affect their microstructural, microchemical, and mechanical characteristics. MATERIALS AND METHODS: In this observational descriptive study, a total of 140 NiTi instruments, 70 ProTaper Gold® (PTG) and 70 WaveOne Gold® (WOG) were analyzed. For each brand system, instruments were evaluated in the as-received condition (n = 10) and after one (n = 20), two (n = 20), and three (n = 20) instrumentation/sterilization cycles. Intraoperative instrumentation parameters were recorded for all used instruments. Afterward, the files were examined using scanning electron microscopy and energy-dispersive X-ray microanalysis. All of the instruments were tensile-fatigue tested until rupture in order to calculate the mechanical tensile strength and the maximum elongation percentage for the samples. Statistical analysis was completed using Chi-square, Kruskal-Wallis H-, or Mann-Whitney U-tests with a statistical significance set at P < 0.05. RESULTS: Significant increasing changes in surface topography (P < 0.05, Chi-square test) and chemical composition (P < 0.05, Kruskal-Wallis H-test) in both brand systems through instrumentation/sterilization cycles were detected. In addition, values of mechanical tensile strength and maximum elongation percentage increased significantly through instrumentation/sterilization cycles in the PTG group, whereas only the median values of mechanical tensile strength increased significantly in the WOG group (all P < 0.01, Kruskal-Wallis H-test). CONCLUSION: Although multiple instrumentation/sterilization cycles may render NiTi instruments more flexible and fatigue resistant, the significant changes detected in their surface topography and chemical composition should preclude their repeated clinical use in the routine endodontic practice as prevention for breakage.

Synthesis of silver nanoparticles using white-rot fungus Anamorphous Bjerkandera sp. R1: influence of silver nitrate concentration and fungus growth time.

Currently, silver nanoparticles (AgNPs) constitute an interesting field of study in medicine, catalysis, optics, among others. For this reason, it has been necessary to develop new methodologies that allow a more efficient production of AgNPs with better antimicrobial and biological properties. In this research growth time effects Anamorphous Bjerkandera sp. R1 and the silver nitrate (AgNO3) concentration over AgNPs synthesis were studied. Through the protocol used in this work, it was found that the action of the capping proteins on the surface of the mycelium played a determining role in the reduction of the Ag+ ion to Ag0 nanoparticles producing a particle size that oscillated between 10 and 100 nm. The progress of the reaction was monitored using visible UV-Vis spectroscopy and the synthesized AgNPs were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Fourier transform infrared radiation (FTIR) spectroscopy. The best synthetic properties were found at 1 mM of AgNO3 concentration, growth time of 8 days, and reaction time of 144 h. Nanometals obtention from microorganisms could be considered as a new method of synthesis, due to reducing abilities of metal ions through its enzymatic system and represents low-cost synthesis that reduces the generation of harmful toxic wastes.

Isolation of chitosan from Ganoderma lucidum mushroom for biomedical applications.

Chitin biopolymer production and its by-product chitosan show great potential. These biomaterials have great applicability in various fields because they are non-toxic, biodegradable, biocompatible, and have antimicrobial effects. The most common source of chitin and chitosan is the crustaceous shell; however, mushrooms are an alternative source for isolating these biopolymers because their cellular wall has a high content of chitin, which may be transformed into chitosan through a deacetylation reaction. The main objective of this research was to obtain chitosan through the deacetylation of chitin isolated from the Ganoderma lucidum basidiomycetes mushroom, which is obtained through biotechnological culture. The material characterization was performed using X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, and an evaluation of cytotoxicity comparing the results obtained with results for commercial chitosan. Protocol results showed that chitosan obtained from this mushroom had a significant similitude with commercial chitosan, yet the one obtained using P2 protocol was the one that rendered the best results: including diffractogram peaks, characteristic infrared analysis bands, and an 80.29 % degree of deacetylation. Cytotoxicity in vitro testing showed that the material was non-toxic; furthermore, it rendered very promising information regarding the evaluation of future applications of this biomaterial in the field of biomedicine.

Comparison of extraction methods of chitin from Ganoderma lucidum mushroom obtained in submerged culture.

The chitin was isolated from the Ganoderma lucidum submerged cultures mycelium as potential source of chitin under biotechnological processes. The extraction of chitin was carried out through 5 different assays which involved mainly three phases: pulverization of the mushroom, deproteinization of the mycelia with NaOH solution, and a process of decolorization with potassium permanganate and oxalic acid. The chitin contents extracted from 9-day mycelia were 413, 339, 87, 78, and 144 mg/g(-1) (milligrams of chitin/grams of dry biomass) for A1, A2, A3, A4, and A5, respectively. Obtained chitin was characterized by X-Ray Diffraction (XRD), by Fourier transform infrared spectroscopy (FTIR), and by thermal analysis (TGA). The results showed that Ganoderma lucidum chitin has similar characteristic of chitin from different fonts. The advantage of the biotechnological processes and the fact that Ganoderma lucidum fungus may be used as a potential raw material for chitin production were demonstrated.