Mineralized alginate hydrogels using marine carbonates for bone tissue engineering applications.

The search for an ideal bone tissue replacement has led to the development of new composite materials designed to simulate the complex inorganic/organic structure of bone. The present work is focused on the development of mineralized calcium alginate hydrogels by the addition of marine derived calcium carbonate biomineral particles. Following a novel approach, we were able to obtain calcium carbonate particles of high purity and complex micro and nanostructure dependent on the source material. Three different types of alginates were selected to develop inorganic/organic scaffolds in order to correlate alginate composition with scaffold properties and cell behavior. The incorporation of calcium carbonates into alginate networks was able to promote extracellular matrix mineralization and osteoblastic differentiation of mesenchymal stem cells when added at 7 mg/ml. We demonstrated that the selection of the alginate type and calcium carbonate origin is crucial to obtain adequate systems for bone tissue engineering as they modulate the mechanical properties and cell differentiation.

Effective genetic modification and differentiation of hMSCs upon controlled release of rAAV vectors using alginate/poloxamer composite systems.

Viral vectors are common tools in gene therapy to deliver foreign therapeutic sequences in a specific target population via their natural cellular entry mechanisms. Incorporating such vectors in implantable systems may provide strong alternatives to conventional gene transfer procedures. The goal of the present study was to generate different hydrogel structures based on alginate (AlgPH155) and poloxamer PF127 as new systems to encapsulate and release recombinant adeno-associated viral (rAAV) vectors. Inclusion of rAAV in such polymeric capsules revealed an influence of the hydrogel composition and crosslinking temperature upon the vector release profiles, with alginate (AlgPH155) structures showing the fastest release profiles early on while over time vector release was more effective from AlgPH155+PF127 [H] capsules crosslinked at a high temperature (50°C). Systems prepared at room temperature (AlgPH155+PF127 [C]) allowed instead to achieve a more controlled release profile. When tested for their ability to target human mesenchymal stem cells, the different systems led to high transduction efficiencies over time and to gene expression levels in the range of those achieved upon direct vector application, especially when using AlgPH155+PF127 [H]. No detrimental effects were reported on either cell viability or on the potential for chondrogenic differentiation. Inclusion of PF127 in the capsules was also capable of delaying undesirable hypertrophic cell differentiation. These findings are of promising value for the further development of viral vector controlled release strategies.

The synergistic effect of VEGF and biomorphic silicon carbides topography on in vivo angiogenesis and human bone marrow derived mesenchymal stem cell differentiation.

Topographical features of biomaterials are able to modulate cell attachment, spreading and differentiation. The addition of growth factors to implantable biomaterials can modify these cellular responses, enhancing their therapeutic potential. The aim of this research is to establish the influence of biomorphic silicon carbide ceramics (bioSiCs) surface topography on the proliferation and osteoblastic differentiation of mesenchymal stem cells and the potential synergistic effect of the ceramic porous structure together with vascular endothelial growth factor loading (VEGF) on the surface mediated osteoblastic differentiation. Three porous bioSiCs with important differences in their microstructure were obtained from different natural precursors. Samples loaded with or without VEGF through ionic interactions were cultured with human umbilical vein endothelial cells (HUVEC) or bone marrow derived mesenchymal stem cells (hMSCs). Cell behaviour and protein activity with regard to bioSiC porous structure and surface properties were analysed. An in vivo model (Chick Chorioallantoic Membrane; CAM) was used to assess the capability of the VEGF loaded systems to promote angiogenesis. Experimental data show that loaded systems were able to control the release of VEGF for up to 15 d ensuring the activity of the protein, increasing the proliferation of HUVECs and the formation of new blood vessels in the CAM. It was found that the selection of bioSiCs with a higher pore size promoted a higher concentration of osteoblastic differentiation markers of MSCs cultured on the surface of bioSiCs. Furthermore, the addition of VEGF to the systems was able to promote a faster osteoblastic differentiation according to the qPCR results, suggesting a synergy between both the surface properties and the controlled release of the growth factor. The VEGF loaded sapelli bioSiC was found to be the most promising material for bone tissue engineering applications.

Controlled release of indomethacin from alginate-poloxamer-silicon carbide composites decrease in-vitro inflammation.

Composites of biomorphic silicon carbides (bioSiCs) and hydrogels are proposed in order to obtain materials able to load and release poor soluble drugs with application in bone pathologies therapy. Hydrogels composed by alginate and poloxamer were loaded with indomethacin, incorporated into the ceramics and crosslinked. The indomethacin release profile is dependent on the microstructure of the bioSiC selected. The loaded oak and sapelli bioSiCs composites have adequate release profiles to promote the decreasing of the secretion of pro-inflammatory cytokines in LPS stimulated macrophages, showing stronger anti-inflammatory effects than pine bioSiC composites. The released indomethacin is able to modulate the degradation of chondrocytes extracellular matrix and promote the formation of new collagen by osteoarthritic chondrocytes. Particles derived from mechanical wear of biomorphic silicon carbides do not show high toxicity, being similar to the zirconia particles.

Key parameters in blood-surface interactions of 3D bioinspired ceramic materials.

Direct contact of materials with blood components may trigger numerous processes which ultimately lead to hemolysis, clot formation and recruitment of inflammatory cells. In this study, the blood-surface interactions for two inert bioinspired ceramic scaffolds obtained from natural resources; biomorphic carbon and silicon carbides (bioSiC) from different origins have been studied. The response of the blood in contact with carbon is well known, however little has been identified on the influence of their 3D porous structure. Moreover, to our knowledge, there is no reference in the literature about the hemocompatibility of biomorphic silicon carbide as a porous scaffold. The experimental results showed the surface energy to be crucial to evaluate the hemocompatibility of a material however the surface topography and material porosity are also parameters to be considered. Surface roughness modifies clot formation whereas for protein adsorption total sample porosity seems to be the key parameter to be considered for hydrophilic materials (biomorphic silicon carbides), while the size of the pores determines the hemolytic response.

Suitability of Biomorphic Silicon Carbide Ceramics as Drug Delivery Systems against Bacterial Biofilms.

The present work is aimed at getting a new insight into biomorphic silicon carbides (bioSiCs) as bone replacement materials. BioSiCs from a variety of precursors were produced, characterized, and loaded with a broad-spectrum antibiotic. The capacity of loaded bioSiCs for preventing and/or treating preformed S. aureus biofilms has been studied. The differences in precursor characteristics are maintained after the ceramic production process. All bioSiCs allow the loading process by capillarity, giving loaded materials with drug release profiles dependent on their microstructure. The amount of antibiotic released in liquid medium during the first six hours depends on bioSiC porosity, but it could exceed the minimum inhibitory concentration of Staphylococcus aureus, for all the materials studied, thus preventing the proliferation of bacteria. Differences in the external surface and the number and size of open external pores of bioSiCs contribute towards the variations in the effect against bacteria when experiments are carried out using solid media. The internal structure and surface properties of all the systems seem to facilitate the therapeutic activity of the antibiotic on the preformed biofilms, reducing the number of viable bacteria present in the biofilm compared to controls.

Smart design of intratumoral thermosensitive ß-lapachone hydrogels by Artificial Neural Networks.

This study presents Artificial Neural Networks (ANN) as a tool for designing injectable intratumoral formulations of the anticancer drug ß-lapachone. This methodology permits insight into the interactions between variables and determines the design space of the formulation without the restrictions of an experimental design. An ANN model for two critical parameters of the formulations; the amount of solubilized drug and gel temperature was developed and validated. The model allowed an understanding of interactions between ingredients in the formulation and the fundamental phenomena as the formation of polypseudorotaxanes to be detected and quantified.

Bio-inspired porous SiC ceramics loaded with vancomycin for preventing MRSA infections.

Implant-related infections are a serious complication in orthopaedic and dental surgery resulting in prolonged hospitalization, high medical costs and patient mortality. The development of porous implants loaded with antibiotics may enable a local delivery for preventing surface colonization and biofilm formation. A new generation of bio-derived porous ceramic material that mimics hierarchical structures from Nature was evaluated. Silicon carbide ceramics derived from Sapelli wood (bioSiC) were obtained by pyrolysis of Entandrophragma cylindricum wood followed by infiltration with molten silicon. This process renders disks that keep the bimodal pore size distribution (3 and 85 µm) of the original material and are highly cytocompatible (BALB/3T3 cell line). The ability of the bio-ceramic to load the antimicrobial agent vancomycin was evaluated by immersion of disks in drug solutions covering a wide range of concentrations. The disks released at pH 7.4 an important amount of drug during the first 2 h (up to 11 mg/g bioSiC) followed by a slower release, which is related to the presence of macro- and mesopores. Finally, the anti-biofilm effect against methicillin resistant Staphylococcus aureus was assessed and a considerable reduction (92%) of the bacterial film was observed. Results highlight the bioSiC potential as component of medicated medical devices.

Inhibition of ciliary activity in organ cultures of ferret trachea in reference to genetic and biological characters in influenza virus strains.

As reported previously, attenuated stable inhibitor-resistant influenza viruses can be screened by a 50% ciliary activity inhibition test in ferret tracheal organ cultures. This test was further applied to a 5 attenuated cold-adapted influenza strains and to 11 strains with known a percentage of RNA-RNA hybridization with the parental A/PR/8/34 (HON1) virus strain. Again, with one exception, attenuated strains could be clearly differentiated from virulent ones. It was concluded that virulence of influenza strains for man can be detected using this test regardless of the techniques used to prepare attenuated variants. A preliminary screening of attenuated candidates for live influenza vaccines can be achieved with confidence on ferret tracheal organ cultures.

Bencilación selectiva del metil 4, 6-0-benciliden-Ó-D-manopiranósido / Selective Benzylation of Methyl 4, 6-0-benzylidene-Ó-d-mannopyranoside

Se intentó llevar a cabo la monobencilación del compuesto (II) siguiendo un método informado en la literatura y se obtuvieron resultados diferentes a los descritos. La estructura de los productos se estableció por hidrólisis suavemente ácida y oxidación peryódica. Además se corroboró ésta por espectrometría de masa, por r. m. n. y por la síntesis de uno de los productos utilizando otro método informado en la literatura. Se ensayaron varios métodos para lograr la obtención selectiva de uno de los dos isómeros monobencilados. Los mejores resultados se obtuvieron utilizando el método de laquilación interfásica. Este compuesto brinda gran interés para la síntesis de oligosacáridos(AU)

In vitro markers for measuring residual virulence for men of live attenuated influenza viruses.

Human and equine influenza strains attenuated by consecutive passages in the presence of normal horse serum (NHS) on allantois-on-shell system (AOS) and administered to human volunteers or horses as possible live virus vaccine candidates were tested in organ cultures of ferret and hamster trachea. Temperature sensitivity, virus replication, interferon induction and neuraminidase activity were also investigated. Observation of the ciliary activity of each ring was made daily. Influenza strains which were incompletely attenuated following two to five passages on AOS + NHS system and caused severe to mild symptoms of influenza in volunteers or horses also caused a 50% inhibition of the activity of the ciliated epithelium earlier than did completely attenuated strains following ten passages on the same system. The A/Hong Kong/68 strain attenuated by Beare and Bynoe was used as a reference strain in every test. This technique appears to be valuable for screening live influenza vaccine candidates and may prevent risking severe illness in volunteers. No significant results could be ascertained with other markers investigated.