Browsing by Author "Monteiro, Fernando J."
Now showing 1 - 4 of 4
Results Per Page
Sort Options
- Development of silk fibroin/nanohydroxyapatite composite hydrogels for bone tissue engineeringPublication . Ribeiro, Marta; Moraes, Mariana A. de; Beppu, Marisa M.; Garcia, Mónica P.; Fernandes, Maria H.; Monteiro, Fernando J.; Ferraz, Maria PiaThis work presents a novel composite hydrogel consisting of silk fibroin (SF) and nanohydroxyapatite (nanoHA) prepared by a new and innovative method using ethanol as gelling agent capable of forming hydrogels in few minutes. The properties of the composite material, such as the microstructure as well as the chemical and physical properties were studied. Moreover in␣vitro studies of osteoblastic citocompatibility were performed. The microporosity and macroporosity obtained combined with interconnected porous structure and a uniform dispersion of nanoHA particles throughout the fibroin matrix makes composite hydrogel suitable for bone regeneration. The compression modulus of composite hydrogels was increased as the nanoHA concentration increased from 10 to 15 wt.% and the water uptake ability of these materials decreased with the incorporation of nanoHA. The metabolic and alkaline phosphatase activities of osteoblastic cells were improved with the incorporation of nanoHA in the SF matrix providing a more promising material for bone tissue engineering.
- Lytic bacteriophages against multidrug-resistant Staphylococcus aureus, Enterococcus faecalis and Escherichia coli isolates from orthopaedic implant-associated infectionsPublication . Barros, Joana; Melo, Luís D.R.; Poeta, Patrícia; Igrejas, Gilberto; Ferraz, Maria Pia; Azeredo, Joana; Monteiro, Fernando J.Orthopaedic implant-associated infections are a devastating complication of orthopaedic surgery with a significant impact on patients and healthcare systems. The aims of this work were to describe the patterns of antimicrobial resistance, pathogenicity and virulence of clinical bacterial isolates from orthopaedic implant-associated infections and to further isolate and characterise bacteriophages that are efficient in controlling these bacteria. Staphylococcus aureus, Enterococcus faecalis and Escherichia coli isolated from orthopaedic infections showed multiresistance patterns to the most frequently used antibiotics in clinical settings. The presence of mobile genetic elements (mecA, Tn916/Tn1545 and intl1) and virulence determinants (icaB, cna, hlb, cylLs, cylM, agg, gelE, fsr and fimA) highlighted the pathogenicity of these isolates. Moreover, the isolates belonged to clonal complexes associated with the acquisition of pathogenicity islands and antimicrobial resistance genes by recombination and horizontal gene transfer. Bacteriophages vB_SauM_LM12, vB_EfaS_LM99 and vB_EcoM_JB75 were characterised and their ability to infect clinical isolates of S. aureus, E. faecalis and E. coli, respectively, was assessed. Morphological and genomic analyses revealed that vB_EfaS_LM99 and vB_EcoM_JB75 belong to the Siphoviridae and Myoviridae families, respectively, and no genes associated with lysogeny were found. The bacteriophages showed low latent periods, high burst sizes, broad host ranges and tolerance to several environmental conditions. Moreover, they showed high efficiency and specificity to infect and reduce clinical bacteria, including methicillin-resistant S. aureus and vancomycin-resistant enterococci. Therefore, the results obtained suggest that the bacteriophages used in this work are a promising approach to control these pathogens involved in orthopaedic implant-associated infections.
- Silk fibroin/nanohydroxyapatite hydrogels for promoted bioactivity and osteoblastic proliferation and differentiation of human bone marrow stromal cellsPublication . Ribeiro, Marta; Fernandes, Maria H.; Beppu, Marisa M.; Monteiro, Fernando J.; Ferraz, Maria PiaSilk fibroin (SF) is a natural, biocompatible, and biodegradable polymer having a great potential for the successful regeneration of damaged bone tissue. In the present work, nanohydroxyapatite (nanoHA) was incorporated into SF polymer to form a bioactive composite hydrogel for applications as bone implants. The degradation and bioactive properties of SF/nanoHA composite hydrogels were evaluated. Additionally, biological investigations of human bone marrow stromal cells (hBMSCs) viability, proliferation and differentiation to the osteoblastic phenotype were conducted. The incorporation of nanoHA in SF polymer matrices improved the bioactivity of the hydrogels. The biological results highlighted that the SF/nanoHA composite hydrogels are suitable for hBMSCs attachment and proliferation, while a test for alkaline phosphatase (ALP) and bone morphogenetic protein 2 (BMP-2) expression suggested osteoblast differentiation. Additionally, a cell staining method for ALP allowed to observe cell infiltration with active production of ALP by the infiltrated cells, paving the way to use the proposed composite hydrogel for bone tissue regeneration.
- Staphylococcus aureusandEscherichia colidual-species biofilms on nanohydroxyapatite loaded with CHX or ZnO nanoparticlesPublication . Barros, Joana; Grenho, Liliana; Fontenente, Sílvia; Manuel, Cândida M.; Nunes, Olga C.; Melo, Luís F.; Monteiro, Fernando J.; Ferraz, Maria PiaImplant-associated infections are caused by surface-adhering microorganisms persisting as biofilms, resistant to host defense and antimicrobial agents. Given the limited efficacy of traditional antibiotics, novel strategies may rely on the prevention of such infections through the design of new biomaterials. In this work, two antimicrobial agents applied to nanohydroxyapatite materials-namely, chlorhexidine digluconate (CHX) and zinc oxide (ZnO) nanoparticles-were compared concerning their ability to avoid single- or dual-species biofilms of Staphylococcus aureus and Escherichia coli. The resulting biofilms were quantified by the enumeration of colony-forming units and examined by confocal microscopy using both Live/Dead staining and bacterial-specific fluorescent in situ hybridization. The sessile population arrangement was also observed by scanning electron microscopy. Both biomaterials showed to be effective in impairing bacterial adhesion and proliferation for either single- or dual-species biofilms. Furthermore, a competitive interaction was observed for dual-species biofilms wherein E. coli exhibited higher proliferative capacity than S. aureus, an inverse behavior from the one observed in single-species biofilms. Therefore, either nanoHA-CHX or nanoHA-ZnO surfaces appear as promising alternatives to antibiotics for the prevention of devices-related infections avoiding the critical risk of antibiotic-resistant strains emergence. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 491-497, 2017.