RESUMO
The growing interest in the use of polysaccharides nanoparticles for biomedical applications is related to the recent progresses on the synthesis of cellulose-based polymers with the specific functionalities. In particular, cellulose graft copolymers are emerging as amphiphilic materials suitable to fabricate smart nanoparticles for drug delivery applications. In this work, a cellulose-graft-poly(ε-caprolactone) (cell-g-PCL) was synthetized and characterized by FTIR, TGA and DSC in order to validate the synthesis process. We demonstrated that fast evaporation processes promoted cell-g-PCL self-assembly to form nanomicellar structures with hydrodynamic radius ranged from 30 to 60â¯nm as confirmed by TEM analysis. Moreover, the application of controlled electrostatic forces on solvent evaporation - namely electrospraying - allowed generating round-like nanoscaled particles, as confirmed by SEM supported via image analysis. We demonstrated also that sodium diclofenac (DS) drastically influenced the mechanism of particle formation, favoring the deposition of erythrocyte-like particles with highly concave surfaces, not penalizing the encapsulation efficiency of nanoparticles (>80%). The release profile showed a fast delivery of DS - about 60% during the first 24â¯h - followed by a sustained release - about 20% during the next 6 days - strictly related to the peculiar weak interactions among amphiphilic polymer segments and water molecules, thus suggesting a successful use of electrosprayed cell-g-PCL nanoparticles for therapeutic treatments in nanomedicine.
Assuntos
Celulose/química , Sistemas de Liberação de Medicamentos/métodos , Nanopartículas/química , Poliésteres/química , Diclofenaco/química , Diclofenaco/farmacocinética , Liberação Controlada de Fármacos , Humanos , Espectroscopia de Ressonância Magnética , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanopartículas/toxicidade , Nanotecnologia/métodos , Polímeros/síntese química , Polímeros/química , Propanóis/química , Solventes/química , Espectroscopia de Infravermelho com Transformada de Fourier , TermogravimetriaRESUMO
This paper reports the experimental assessment of an automated optical assay based on label free optical fiber optrodes for the fast detection of class C ß-lactamases (AmpC BLs), actually considered as one of the most important sources of resistance to ß-lactams antibiotics expressed by resistant bacteria. Reflection-type long period fiber gratings (RT-LPG) have been used as highly sensitive label free optrodes, while a higher affine boronic acid-based ligand was here selected to enhance the overall assay performances compared to those obtained in our first demonstration. In order to prove the feasibility analysis towards a fully automated optical assay, an engineered system was developed to simultaneously manipulate and interrogate multiple fiber optic optrodes in the different phases of the assay. The automated system tested in AmpC solutions at increasing concentrations demonstrated a limit of detection (LOD) of 6 nM, three times better when compared with the results obtained in our previous work. Moreover, the real effectiveness of the proposed optical assay has been also confirmed in complex matrices as the case of lysates of Escherichia coli overexpressing AmpC.
RESUMO
We report an innovative fiber optic nano-optrode based on Long Period Gratings (LPGs) working in reflection mode for the detection of human Thyroglobulin (TG), a protein marker of differentiated thyroid cancer. The reflection-type LPG (RT-LPG) biosensor, coated with a single layer of atactic polystyrene (aPS) onto which a specific, high affinity anti-Tg antibody was adsorbed, allowed the label-free detection of Tg in the needle washouts of fine-needle aspiration biopsies, at concentrations useful for pre- and post-operative assessment of the biomarker levels. Analyte recognition and capture were confirmed with a parallel on fiber ELISA-like assay using, in pilot tests, the biotinylated protein and HRP-labeled streptavidin for its detection. Dose-dependent experiments showed that the detection is linearly dependent on concentration within the range between 0 and 4 ng/mL, while antibody saturation occurs for higher protein levels. The system is characterized by a very high sensitivity and specificity allowing the ex-vivo detection of sub ng/ml concentrations of human Tg from needle washouts of fine-needle aspiration biopsies of thyroid nodule from different patients.