In situ preparation and protein delivery of silicate–alginate composite microspheres with core-shell structure

The efficient loading and sustained release of proteins from bioactive microspheres remain a significant challenge. In this study, we have developed bioactive microspheres which can be loaded with protein and then have a controlled rate of protein release into a surrounding medium. This was achieved by preparing a bioactive microsphere system with core-shell structure, combining a calcium silicate (CS) shell with an alginate (A) core by a one-step in situ method. The result was to improve the microspheres'' protein adsorption and release, which yielded a highly bioactive material with potential uses in bone repair applications. The composition and the core-shell structure, as well as the formation mechanism of the obtained CS–A microspheres, were investigated by X-ray diffraction, optical microscopy, scanning electron microscopy, energy dispersive spectrometer dot and line-scanning analysis. The protein loading efficiency reached 75 per cent in CS–A microspheres with a core-shell structure by the in situ method. This is significantly higher than that of pure A or CS–A microspheres prepared by non-in situ method, which lack a core-shell structure. CS–A microspheres with a core-shell structure showed a significant decrease in the burst release of proteins, maintaining sustained release profile in phosphate-buffered saline (PBS) at both pH 7.4 and 4.3, compared with the controls. The protein release from CS–A microspheres is predominantly controlled by a Fickian diffusion mechanism. The CS–A microspheres with a core-shell structure were shown to have improved apatite-mineralization in simulated body fluids compared with the controls, most probably owing to the existence of bioactive CS shell on the surface of the microspheres. Our results indicate that the core-shell structure of CS–A microspheres play an important role in enhancing protein delivery and mineralization, which makes these composite materials promising candidates for application in bone tissue regeneration.     

Relative larvicidal property of common oxide nanostructures against Culex quinquefasciatus

Common oxide nanostructures such as silicon‐di‐oxide, magnesium‐oxide, zinc‐oxide, and copper‐oxide (CuO) having useful functional and bioactive properties have been synthesised and characterised. All these nanostructures have been found to be larvicidal towards Culex quinquefasciatus mosquito especially against lower instars in comparison with higher instars in 48 h. Only, CuO is larvicidal against late instar stages after 48 h. Moreover, CuO is larvicidal against first instar stages after 24 h (LC₅₀ 157 mg/l). However, none of these nanostructures are pupicidal. Post mortality larval morphology was found to be distorted under bright field microscopy and scanning electron microscopy images of affected larval surface appeared to be rough and uneven. Fluorescent images showed that nanostructures infiltrated inside visceral organs of larvae. Nanostructures also caused tissue oxidative stress in larvae. These results indicate that above stated oxide nanostructures are effective larvicidal agents against early instar stages of Culex larvae.Inspec keywords: biodegradable materials, oxidation, optical microscopy, fluorescence, cellular biophysics, biological tissues, diseases, scanning electron microscopy, toxicology, biological techniquesOther keywords: late instar stages, bright field microscopy, scanning electron microscopy images, tissue oxidative stress, oxide nanostructures, silicon‐dioxide, magnesium‐oxide, zinc‐oxide, copper‐oxide, functional properties, bioactive properties, larvicidal property, larvicidal agents, Culex quinquefasciatus mosquito, Culex larvae     

Production,stabilisation and characterisation of silver nanoparticles coated with bioactive polymers pluronic F68, PVP and PVA

The increasing and alarming panorama of bacterial infections and associated morbidities that occur during medical and hospital procedures makes the development of technologies that aid in controlling such bacterial infections of utmost importance. Recent studies have shown that formulations with metal nanoparticles exhibit good antibacterial properties against a broad spectrum of microorganisms. Moreover, it was demonstrated that some biologically active polymeric materials, when applied in combination with chemical antimicrobial agents, enhance the therapeutic action of the latter. The research effort entertained herein aimed at the physico‐chemical characterisation of silver nanoparticles obtained by chemical reduction, stabilised by bioactive polymers polyvinyl alcohol and polyvinylpyrrolidone, and further co‐stabilised by pluronic F68. Scanning electron microscopy images of the nanoparticles produced, coated with different stabilisers, have shown that the chemical nature of the stabilisation effect promoted incorporation of pluronic in the nanoparticles and was closely related to an increase in the silver concentration in the nanoparticle samples obtained via energy‐dispersive X‐ray spectroscopy. The study described herein also shows that the nature of the stabiliser favours the interaction of pluronic F68 with samples containing silver nanoparticles.Inspec keywords: silver, nanoparticles, polymer films, coatings, nanocomposites, nanofabrication, microorganisms, biomedical materials, nanomedicine, antibacterial activity, reduction (chemical), scanning electron microscopy, X‐ray chemical analysisOther keywords: bioactive polymers pluronic F68 coated silver nanoparticles, PVP coated silver nanoparticles, PVA coated silver nanoparticles, bacterial infections, associated morbidities, medical procedures, hospital procedures, antibacterial properties, microorganisms, biologically active polymeric materials, chemical antimicrobial agents, therapeutic action, physicochemical characterisation, chemical reduction, bioactive polymers polyvinyl alcohol, polyvinylpyrrolidone, scanning electron microscopy, stabilisation effect, energy‐dispersive X‐ray spectroscopy, Ag     

Effect of nanoparticulate bioactive glass particles on bioactivity and cytocompatibility of poly(3-hydroxybutyrate) composites

This work investigated the effect of adding nanoparticulate (29 nm) bioactive glass particles on the bioactivity, degradation and in vitro cytocompatibility of poly(3-hydroxybutyrate) (P(3HB)) composites/nano-sized bioactive glass (n-BG). Two different concentrations (10 and 20 wt %) of nanoscale bioactive glass particles of 45S5 Bioglass composition were used to prepare composite films. Several techniques (Raman spectroscopy, scanning electron microscopy, atomic force microscopy, energy dispersive X-ray) were used to monitor their surface and bioreactivity over a 45-day period of immersion in simulated body fluid (SBF). All results suggested the P(3HB)/n-BG composites to be highly bioactive, confirmed by the formation of hydroxyapatite on material surfaces upon immersion in SBF. The weight loss and water uptake were found to increase on increasing bioactive glass content. Cytocompatibility study (cell proliferation, cell attachment, alkaline phosphatase activity and osteocalcin production) using human MG-63 osteoblast-like cells in osteogenic and non-osteogenic medium showed that the composite substrates are suitable for cell attachment, proliferation and differentiation.     

In vitro investigation of novel calcium phosphates using osteogenic cultures

A rat bone marrow stromal cell (RBM) culture was used to evaluate novel bioactive calcium phosphate ceramics. Three rapidly resorbable, glassy crystalline materials with the main crystalline phase Ca2KNa(PO4)2 were investigated (sample code GB 1a, GB 14, GB 9). These materials were designed to exhibit a higher degree of biodegradability than tricalcium phosphate. Additionally, a bioactive glass ceramic of low biodegradability was examined (sample code AP 40). RBM cells were cultured on the disc-shaped test substrata for 14 d. The culture medium was changed and calcium and phosphate concentrations of the medium were determined daily. Specimens were evaluated using light microscopy and morphometry of the cell-covered substrate surface, scanning electron microscopy and energy dispersive X-ray analysis. Except for GB 1a, the rat bone marrow cells attached and grew on all substrate surfaces. Of the different calcium phosphate ceramics tested, AP 40 facilitated osteoblast growth and the elaboration of the extracellular matrix to the highest degree followed by GB 9 and GB 14. The inhibition of cell growth encountered with GB 1a seemed to be related to its high phosphate ion release. © 1998 Chapman & Hall     

From tissue retrieval to electron tomography: nanoscale characterization of the interface between bone and bioactive glass

The success of biomaterials for bone regeneration relies on many factors, among which osseointegration plays a key role. Biogran (BG) is a bioactive glass commonly employed as a bone graft in dental procedures. Despite its use in clinical practice, the capability of BG to promote osseointegration has never been resolved at the nanoscale. In this paper, we present the workflow for characterizing the interface between newly formed bone and BG in a preclinical rat model. Areas of bone–BG contact were first identified by backscattered electron imaging in a scanning electron microscope. A focused ion beam in situ lift-out protocol was employed to prepare ultrathin samples for transmission electron microscopy analysis. The bone–BG gradual interface, i.e. the biointerphase, was visualized at the nanoscale with unprecedented resolution thanks to scanning transmission electron microscopy. Finally, we present a method to view the bone–BG interface in three dimensions using electron tomography.     

Synthesis and characterisation of PEG modified chitosan nanocapsules loaded with thymoquinone

Thymoquinone (TQ), a major bioactive compound of Nigella sativa seeds has several therapeutic properties. The main drawback in bringing TQ to therapeutic application is that it has poor stability and bioavailability. Hence a suitable carrier is essential for TQ delivery. Recent studies indicate biodegradable polymers are potentially good carriers of bioactive compounds. In this study, polyethylene glycol (PEG) modified chitosan (Cs) nanocapsules were developed as a carrier for TQ. Aqueous soluble low molecular weight Cs and PEG was selected among different biodegradable polymers based on their biocompatibility and efficacy as a carrier. Optimisation of synthesis of nanocapsules was done based on particle size, PDI, encapsulation efficiency and process yield. A positive zeta potential value of +48 mV, indicating good stability was observed. Scanning electron microscope and atomic‐force microscopy analysis revealed spherical shaped and smooth surfaced nanocapsules with size between 100 to 300 nm. The molecular dispersion of the TQ in Cs PEG nanocapsules was studied using X‐ray powder diffraction. The Fourier transform infrared spectrum of optimised nanocapsule exhibited functional groups of both polymer and drug, confirming the presence of Cs, PEG and TQ. In vitro drug release studies showed that PEG modified Cs nanocapsules loaded with TQ had a slow and sustained release.Inspec keywords: nanomedicine, drug delivery systems, polymers, scanning electron microscopy, electrokinetic effects, atomic force microscopy, X‐ray diffraction, Fourier transform infrared spectraOther keywords: PEG modified chitosan nanocapsules, thymoquinone, bioactive compound, Nigella sativa seeds, bioavailability, polyethylene glycol, molecular weight, zeta potential, scanning electron microscope, atomic force microscopy, molecular dispersion, X‐ray powder diffraction, Fourier transform infrared spectrum     

Coated biodegradable casein nanospheres: a valuable tool for oral drug delivery

Abstract

Biodegradable casein nanospheres for the sustained release of bioactive molecules in the gastro-intestinal tract were prepared by precipitation polymerization using sodium methacrylate (NaMA) and N,N′-methylene bis-acrylamide (MEBA) as pH-responsive monomer and cross-linker. Three materials with different casein amount were obtained and characterized by scanning electron microscopy, dimensional analysis, water uptake, cytotoxicity and enzymatic degradation experiments. Nanospheres biodegradability was tuned by coating with polyacrylic acid. Coated and uncoated materials were investigated as delivery vehicles for diclofenac sodium salt. For un-coated samples, the release raise 100% in 30?h, while for coated specimens these values were lower than 70%, due to the diffusional constraints of polymer layer.     

Fabrication of nanocrystalline hydroxyapatite doped degradable composite hollow fiber for guided and biomimetic bone tissue engineering

Natural bone tissue possesses a nanocomposite structure interwoven in a three-dimensional (3-D) matrix, which plays critical roles in conferring appropriate physical and biological properties to the bone tissue. Single type of material may not be sufficient to mimic the composition, structure and properties of native bone, therefore, composite materials consisting of both polymers, bioceramics, and other inorganic materials have to be designed. Among a variety of candidate materials, polymer-nanoparticle composites appear most promising for bone tissue engineering applications because of superior mechanical properties, improved durability, and surface bioactivity when compared with conventional polymers or composites. The long term objective of this project is to use highly aligned, bioactive, biodegradable scaffold mimicking natural histological structure of human long bone, and to engineer and regenerate human long bone both in vitro and in vivo. In this study, bioactive, degradable, and highly permeable composite hollow fiber membranes (HFMs) were fabricated using a wet phase phase-inversion approach. The structure of the hollow fiber membranes was examined using scanning electron microscopy (SEM); degradation behavior was examined using weigh loss assay, gel permeation chromatography (GPC), and differential scanning calorimetry (DSC); and bioactivity was evaluated with the amount of calcium deposition from the culture media onto HFM surface. Doping PLGA HFMs with nanoHA results in a more bioactive and slower degrading HFM than pure PLGA HFMs.     

An innovative auto-catalytic deposition route to produce calcium–phosphate coatings on polymeric biomaterials

The aim of this research is to develop a new methodology to obtain bioactive coatings on bioinert and biodegradable polymers that are not intrinsically bioactive. In this study three types of materials were used as substrates: (i) high molecular weight polyethylene (HMWPE) and two different types of starch based blends (ii) starch/ethylene vinyl alcohol blends, SEVA-C and (iii) starch/cellulose acetate blends, SCA. Two types of baths were originally proposed and studied to produce novel auto-catalytic calcium–phosphate (Ca–P) coatings. Then, the coated surfaces were analyzed by scanning electron microscopy and energy dispersive spectroscopy (SEM/EDS), as produced, and after different immersion periods in SBF. The evolution of Ca and P concentrations was determined by induced-coupled plasma emission (ICP) spectroscopy. The crystalline phases present on the films formed on the different material surfaces, after a certain soaking time, were identified by thin-film X-ray diffraction (TF-XRD). The obtained results indicated that it was possible to coat the materials surfaces with a Ca–P layer with only 60 min of immersion in both types of auto-catalytic solutions. Furthermore, it was possible to observe the clear bioactive nature of the Ca–P coatings after different immersion periods in a simulated body fluid (SBF). The results from TF-XRD confirmed the presence of partially amorphous Ca–P films with clearly noticeable hydroxylapatite peaks. These new methodologies allow for the production of an adherent bioactive film on the polymeric surfaces prior to implantation, which may allow for the development of bone-bonding, bioabsorbable implants and fixation devices. ©2003 Kluwer Academic Publisher     

Silver-containing mesoporous bioactive glass with improved antibacterial properties

The aim of the present work is the study of the bacteriostatic/bactericidal effect of a silver-containing mesoporous bioactive glass obtained by evaporation-induced self-assembly and successive thermal stabilization. Samples of the manufactured mesophase were characterized by means of transmission electron microscopy and N₂ adsorption/desorption at 77 K, revealing structural and textural properties similar to SBA-15 mesoporous silica. Glass samples used for bioactivity experiments were put in contact with a standardized, commercially available cell culture medium instead of lab-produced simulated body fluid, and were then characterized by means of X-ray diffraction, field emission scanning electron microscopy and Fourier transform infrared spectroscopy. All these analyses confirmed the development of a hydroxyl carbonate apatite layer on glass particles. Moreover, the investigated mesostructure showed a very good antibacterial effect against S. aureus strain, with a strong evidence of bactericidal activity already registered at 0.5 mg/mL of glass concentration. A hypothesis about the mechanism by which Ag affects the bacterial viability, based on the intermediate formation of crystalline AgCl, was also taken into account. With respect to what already reported in the literature, these findings claim a deeper insight into the possible use of silver-containing bioactive glasses as multifunctional ceramic coatings for orthopedic devices.     

Tip preparation for usage in an ultra-low temperature UHV scanning tunneling microscope

This work deals with the preparation and characterization of tungsten tips for the use in UHV low-temperature scanning tunneling microscopy and spectroscopy (STM and STS, respectively). These specific environments require in situ facilities for tip conditioning, for further sharpening of the tips, as well as for reliable tip characterization. The implemented conditioning methods include direct resistive annealing, annealing by electron bombardment, and self-sputtering with noble gas ions. Moreover, results from in situ tip characterization by field emission and STM experiments were compared to ex situ scanning electron microscopy. Using the so-prepared tips, high resolution STM images and tunneling spectra were obtained in a temperature range from ambient down to 350 mK, partially with applied magnetic field, on a variety of materials.     

Feasibility, tailoring and properties of polyurethane/bioactive glass composite scaffolds for tissue engineering

This research work aims to propose highly porous polymer/bioactive glass composites as potential scaffolds for hard-tissue and soft-tissue engineering. The scaffolds were prepared by impregnating an open-cells polyurethane sponge with melt-derived particles of a bioactive glass belonging to the SiO₂–P₂O₅–CaO–MgO–Na₂O–K₂O system (CEL2). Both the starting materials and the composite scaffolds were investigated from a morphological and structural viewpoint by X-ray diffraction analysis and scanning electron microscopy. Tensile mechanical tests, carried out according to international ISO and ASTM standards, were performed by using properly tailored specimens. In vitro tests by soaking the scaffolds in simulated body fluid (SBF) were also carried out to assess the bioactivity of the porous composites. It was found that the composite scaffolds were highly bioactive as after 7 days of soaking in SBF a HA layer grew on their surface. The obtained polyurethane/CEL2 composite scaffolds are promising candidates for tissue engineering applications.     

Influence of strontium for calcium substitution on the glass–ceramic network and biomimetic behavior in the ternary system SiO<Subscript>2</Subscript>–CaO–MgO

Strontium (Sr) enhances bone formation both in vitro and in vivo, while it reduces bone resorption. Thus, Sr incorporation in bioactive glass–ceramic scaffolds for bone tissue regeneration could further enhance osteogenesis. The aim of this work was the synthesis, characterization and investigation of the apatite-forming ability in inorganic environment of two sol–gel-derived bioactive Sr-containing glass–ceramic materials with 5 and 10% of SrO. The thermal properties of the synthesized materials were studied using differential thermal analysis (TG–DTA). The apatite-forming ability test was conducted in SBF for various immersion times for both thermally treated and untreated samples. The characterization of the samples before and after immersion in SBF was performed with Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD) and scanning electron microscopy with associated energy-dispersive spectroscopy. FTIR spectra revealed that all synthesized glass–ceramic materials presented the characteristic bands of silicate glasses, while XRD identified various crystalline phases, mostly calcium silicates. Strontium is present in the form of strontium silicate in both as-received and thermally treated specimens, and Sr-diopside in the thermally treated specimens. The apatite-forming ability of the glass–ceramic materials was confirmed by the formation of a hydroxyapatite layer after 3 and 5 days of immersion in SBF on the surface of the untreated and thermally treated samples, respectively. The apatite layer, also, became thicker as the immersion time increased.     

Designing antimicrobial bioactive glass materials with embedded metal ions synthesized by the sol–gel method

Bioactive glasses (SiO₂–P₂O₅–CaO) having tailored concentrations of different biocide metal ions (copper or silver) were produced by the sol–gel method. All the particles release phosphorous ions when immersed in water and simulated body fluid (SBF). Moreover, a surface layer of polycrystalline hydroxy-carbonate apatite was formed on the particle surfaces after 10 day immersion in SBF as confirmed by X-ray diffraction and scanning electron microscopy (SEM) showing the bioactive materials. Samples with embedded either copper or silver ions were able to further release the biocide ions with a release rate that depends on the metal embedded and the dissolution medium: water or SBF. This biocide ion release from the samples explains the antimicrobial effect of our active particles against Escherichia coli DH5α ampicillin-resistant (Gram-negative) and Streptococcus mutans (Gram-positive) as determined by the Minimum Bactericidal Concentration (MBC) method. The antimicrobial behavior of the particles depends on the bacteria and the biocide ion used. Noteworthy, although samples with copper are able to release more metal ion than samples with silver, they present higher MBC showing the high effect of silver against these bacteria.     

Green polyurethane nanocomposites from soy polyol and bacterial cellulose

With increased environmental concerns, fluctuations in oil prices, and dependency on oil, there has been an emergence in the use of biobased polyurethanes prepared with polyols derived from plant oils, such as soybean oil. In this study, novel polyurethane materials were synthesized using polyols obtained from soybean oils. The polyurethanes were produced by reacting the polyols with polymeric isocyanate with an isocyanate index of 100 at 150 °C for 2 h for complete curing. The mechanical properties of this biobased polyurethane were improved by incorporating novel nanosized cellulose produced from bacteria. The source of the bacterial cellulose nanofibrils was a commercially available food product nata-de-coco. A fine dispersion of the nanocellulose fibrils in biobased polyurethane matrix was achieved by using a high speed homogenizer at 30,000 rpm, which was observed by field emission transmission electron microscopy and scanning probe microscopy. The average diameter size of the cellulose fibers were determined to be 22 ± 5 nm by scanning probe microscopy. The flexural strength and modulus were improved even at 0.125 wt% bacterial cellulose concentration and the optimum nanocomposite was obtained with 0.250 wt% concentration due to good interaction of isocyanates and the cellulose. Dynamic mechanical analysis supported the flexural testing data for modulus values. Transparent thick nanocomposite samples show one additional advantage of the nanocomposite technology.     

Precipitation of calcium phosphate in the presence of albumin on titanium implants with four different possibly bioactive surface preparations. An in vitro study

The aim of the present study was to compare the nucleating behaviour on four types of bioactive surfaces by using the simulated body fluid (SBF) model with the presence albumin. Titanium discs were blasted (B) and then prepared by alkali and heat treatment (AH), anodic oxidation (AO), fluoridation (F), or hydroxyapatite coating (HA). The discs were immersed in SBF with 4.5 mg/ml albumin for 3 days, 1, 2, 3 and 4 weeks and analysed with scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX) and X-ray photoelectron spectroscopy (XPS). Topographic surface characterisation was performed with a contact stylus profilometer. The results demonstrated that the bioactive surfaces initiated an enhanced calcium phosphate (CaP) formation and a more rapid increase of protein content was present on the bioactive surfaces compared to the blasted control surface. The observation was present on all bioactive surfaces. The fact that there was a difference between the bioactive surfaces and the blasted control surface with respect to precipitation of CaP and protein content on the surfaces support the fact that there may be biochemical advantages in vivo by using a bioactive surface.     

Preparation and in vitro bioactivity of poly(d,l-lactide) composite containing hydroxyapatite nanocrystals

The nano-sized hydroxyapatite (n-HA) was incorporated into poly(d,l-Lactide) (PDLLA) to form a bioactive and biodegradable composite for application in hard tissue replacement and regeneration. Thin film of PDLLA composite containing 20 mass% of n-HA fillers was successfully developed through integration of solvent co-blending and hot pressing techniques. firstly, n-HA and PDLLA were chemically synthesized, respectively, then mixed together and homogeneously dispersed in N,N-dimethyl formamide(DMF) solvent, finally, the dried blended hybrid containing PDLLA matrix and n-HA fillers was put into the mould and compacted by hot-pressing machine under 8 MPa pressure at 110 °C for 15 min. In vitro studies were conducted using the simulated body fluid(SBF). Composite specimens were soaked in SBF from 1 day to 21 days prior to surface analysis. Results obtained from scanning electron microscopy(SEM) examination, Energy dispersive X-ray detector(EDX) analysis and X-ray diffraction (XRD) analysis showed that a layer of non-stoichiometric apatite formed within 7 days on HA/PDLLA composite surface after its immersion in SBF, demonstrating moderate in vitro bioactivity of n-HA/PDLLA composite, though a moderate rate of apatite formation in SBF was found on initial stage of immersion periods for n-HA/PDLLA composite, compared to the other biomaterial composite. This type of composite film exhibited certain desirable bioactive characteristics, and they are promising bone candidates to develop novel bioactive composites for biomedical application.     

Preparation and thermal stability of amine-terminated polyesterimide modified epoxy resin

In this research, a novel amine-terminated polyesterimide (AT-PEsI) having 1.324 Pa.s of inherent viscosity was synthesized from polyester and pyromellitic dianhydride, p-toluene sulphonic acid as catalyst and characterized by FT-IR and ¹H-NMR spectroscopy. Epoxy resin was modified by the soluble and fusible AT-PEsI prepolymer for improving both thermal and mechanical properties of electronic packaging materials. The AT-PEsI modified epoxy resin was cured by 4,4-diaminodiphenyl ether. The thermal stability of the cured materials was studied with thermogravimetric analysis (TGA) and thermo-mechanical analysis (TMA). The coefficients of thermal expansion of the modified ER materials decrease with the content of AT-PEsI in ER. The results from TMA and TGA show that modified epoxy resins have better thermal stability than pure ER. The morphology of modified ER materials was also evaluated by scanning electron microscopy for the investigation of fracture surface. The results show that the AT-PEsI modified epoxy resin materials exhibit better toughness than pure ER.