ABSTRACT
Bioglass nanoparticles (n-BGs, 54SiO2 :40CaO:6P2 O5 mol %) with about 27 nm diameter were synthesized by the sol-gel method and incorporated into a poly(lactic acid) (PLA) matrix by the melting process in order to obtain nanocomposites with filler contents of 5, 10, and 25 wt %. Our results showed that during the cooling scan, the crystallization temperature (Tc ) of the PLA/n-BG nanocomposites decreased 13°C as compared to neat PLA. The presence of nanoparticles also decreased the thermal stability of the PLA matrix, as nanocomposites presented up to about 20°C lower degradation temperatures in a nitrogen atmosphere. The presence of n-BG increased the stiffness of the polymer matrix, and for instance the composite with 25 wt % of filler presented about 52.6% higher Young's modulus than neat PLA. n-BG incorporation into PLA increased also the hydrolytic degradation of the polymer over time. When the PLA composites were immersed in simulated body fluid, an apatite layer was formed on their surface, as verified by Fourier transform infrared, X-Ray Diffraction (XRD), and scanning electron microscopy-EDS, showing that the presence of n-BG induced bioactivity on the PLA matrix. Moreover, the viability of cervical uterine adenocarcinoma cells was higher on PLA/n-BG nanocomposite with 25 wt % of filler. The presence of n-BG barely gave an antibacterial effect on the polymer matrix, despite the well-known biocidal properties of these nanoparticles. Our results show that the presence of n-BGs is a proper route for improving the bioactivity of PLA with potential application in tissue engineering.
Subject(s)
Biocompatible Materials/chemistry , Ceramics/chemistry , Nanoparticles/chemistry , Polyesters/chemistry , Biocompatible Materials/pharmacology , Cell Survival/drug effects , Ceramics/pharmacology , Crystallization , Elastic Modulus , HeLa Cells , Humans , Nanocomposites/chemistry , Polyesters/pharmacologyABSTRACT
Un paso crucial en el desarrollo de un inmunosensor piezoeléctrico para la detección de tuberculosis (TB), es la selección y obtención de los inmunoreactivos empleados en el inmunoensayo y la estrategia para la biofuncionalización del transductor. Diversos estudios han reportado el uso del antígeno proteico 38kDa (Ag38kDa) de Mycobacterium tuberculosis (Mtb) como un buen biomarcador de la enfermedad y el cumplimiento de las características físicas y bioquímicas para ser inmovilizado por monocapas autoensambladas (SAMs), en la superficie del electrodo de oro de cristales piezoeléctricos. Un inmunosensor piezoeléctrico desarrollado a partir de un antígeno nativo purificado de Mtb podría ser un método alternativo simple para la detección de Mtb con ventajas de rapidez y reusabilidad, contribuyendo al control y el tratamiento oportuno de la enfermedad. En este estudio se presenta el proceso de purificación del Ag38kDa a partir de proteínas de secreción filtradas de cultivo (CFP) de Mtb para ser usado como inmunoreactivo con potencial aplicación en la detección de Mtb con inmunosensores piezoeléctricos. Se obtuvieron cristales funcionalizados mediante la técnica modificada de monocapas autoensambladas (SAMs), con el antígeno nativo purificado y CFP. Las superficies biofuncionalizadas fueron caracterizadas cualitativamente con microscopía de fuerza atómica (AFM) para validar las condiciones de optimización del protocolo de inmovilización con antígenos de secreción de Mtb. Estos cristales modificados pueden ser acoplados a un sistema de caracterización de un inmunosensor piezoeléctrico para la detección de Mtb mediante un inmunoensayo competitivo directo.
The selection and procurance of the immunoreagents used in the immunoassay and biofunctionalisation transducer strategy, are a key in the piezoelectric immunosensor development for the detection of tuberculosis (TB). Many have reported the use of 38kDa protein antigen (Ag38kDa) from Mycobacterium tuberculosis (Mtb) such as good biomarker of TB disease and compliance with physical and biochemical characteristics to be immobilized by self-assembled monolayers (SAMs), in the gold electrode of piezoelectrics crystals surfaces. A piezoelectric immunosensor developed from purified native antigens of Mtb may be an alternative simple method for detection of Mtb with speed and reusable advantages, contributing to the control and early treatment of disease. In this paper, the purification process of Ag38kDa Mtb from secretory proteins filtered culture (CFP) from Mtb is presented as an immunoreactive with potential application in the detection of Mtb by piezoelectric immunosensors. Functionalized crystals were obtained by using the modified self-assembled monolayers (SAMs) technique, with purified native antigen and CFP. The functionalized surfaces were qualitatively characterized using atomic force microscopy (AFM) in order to validate the immobilization protocol optimal conditions for secretion antigens from Mtb. These modified crystals may be coupled to piezoelectric immunosensor characterization system for detecting of Mtb by a direct competition immunoassay.