RESUMO
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.
Assuntos
Materiais Biocompatíveis , Quitosana/isolamento & purificação , Reishi/química , Acetilação , Animais , Biomassa , Linhagem Celular , Quitosana/química , Camundongos , Espectroscopia de Infravermelho com Transformada de Fourier , Termogravimetria , Difração de Raios XRESUMO
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.
Assuntos
Biotecnologia , Técnicas de Cultura de Células , Quitina/química , Quitina/isolamento & purificação , Biomassa , Reatores Biológicos , Reishi/química , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
El quitosano está presente en el caparazón de los crustáceos, y desde hace algún tiempo ha sido utilizado en el campo de la medicina y la ingeniería de tejidos para la fabricación de matrices de crecimiento celular. En este estudio se extrajo quitosano de caparazón de crustáceos y se propuso un método sencillo para fabricar matrices con microestructura controlada. Las matrices fueron preparadas por congelación y liofilización de soluciones de quitosano y luego fueron caracterizadas por microscopía electrónica de barrido. La difracción de rayos X del quitosano extraído mostró un espectro acorde con una fuente comercial del material, evidenciando la efectividad del protocolo de extracción. La microscopía mostró poros ovalados y circulares distribuidos en todo el volumen de las muestras, con diámetros de poros entre 100 µm y 150 µm. Lo anterior demuestra que el método de producción propuesto proporciona un punto de partida para la fabricación de matrices de crecimiento celular.
Chitosan is present in crustacean shells and it has been used in the fields of medicine and tissue engineering for the construction of scaffolds that support cell growth. In this study, chitosan was extracted from crustacean shells and processed into scaffolds with controlled microstructure using a simple processing method presented herein. The scaffolds were prepared by freezing and lyophilization of chitosan solutions and were characterized by scanning electron microscopy. The results showed a chitosan with an X-ray diffraction spectrum similar to that of a commercial chitosan, thus demonstrating the effectiveness of the extraction protocol. Microscopy showed oval and circular pores distributed on the bulk sample, with pore diameters between 100 µm and 150 µm. This shows that the proposed fabrication method provides a starting point for the construction of porous scaffolds that may support cell growth.