Optimized Electrophoretic Deposition of Chitosan/Mesoporous Glass Nanoparticles with Gentamicin on Titanium Implants: Enhancing Hemocompatibility and Antibacterial Activity.

Titanium-based implants have long been studied and used for applications in bone tissue engineering, thanks to their outstanding mechanical properties and appropriate biocompatibility. However, many implants struggle with osseointegration and attachment and can be vulnerable to the development of infections. In this work, we have developed a composite coating via electrophoretic deposition, which is both bioactive and antibacterial. Mesoporous bioactive glass particles with gentamicin were electrophoretically deposited onto a titanium substrate. In order to validate the hypothesis that the quantity of particles in the coatings is sufficiently high and uniform in each deposition process, an easy-to-use image processing algorithm was designed to minimize human dependence and ensure reproducible results. The addition of loaded mesoporous particles did not affect the good adhesion of the coating to the substrate although roughness was clearly enhanced. After 7 days of immersion, the composite coatings were almost dissolved and released, but phosphate-related compounds started to nucleate at the surface. With a simple and low-cost technique like electrophoretic deposition, and optimized stir and suspension times, we were able to synthesize a hemocompatible coating that significantly improves the antibacterial activity when compared to the bare substrate for both Gram-positive and Gram-negative bacteria.

In situ drug release measuring in α-TCP cement by electrochemical impedance spectroscopy.

The use of drug delivery systems is a good technique to leave the right quantity of medicine in the patient's body in a suitable dose, because the drug application is delivered directly to the affected region. The current techniques such as HPLC and UV-Vis for the drug delivery calculation has some disadvantages, as the accuracy and the loss of the sample after characterization. With the aim of reducing the amount of material used during the characterization and have a non-destructive test with instantaneous results, the present paper shows the possibility of using electrochemical impedance spectroscopy (EIS) to have a drug delivery measurement during the release phenomena for a calcium phosphate cement (CFC) delivery system with gentamicin sulfate (GS) and lidocaine hydrochloride (LH), at a ratio of 1% and 2%, respectively. The equivalent circuit and the chemical mechanism involved during the measurements have been proposed as a tool to determine the drug delivery profile. The method has been compared with the UV-Vis technique. XRD was realized to verify conditions, before and after release. It was possible to verify the potential for using EIS as an instant technique to quantify drug delivery.

Dioclea violacea lectin modulates the gentamicin activity against multi-resistant strains and induces nefroprotection during antibiotic exposure.

Gentamicin is an aminoglycoside antibiotic used to treat infections of various origins. In the last few decades, the constant use of gentamicin has resulted in increased bacterial resistance and nephrotoxicity in some cases. In this study, we examined the ability of Dioclea violacea lectin (DVL) in modulate the antimicrobial activity of gentamicin and reduce the nephrotoxicity induced by this drug. The minimum inhibitory concentration (MIC) obtained for DVL against all strains studied was not clinically relevant (MIC ≥ 1024 µg/mL). However, when DVL was combined with gentamicin, a significant increase in antibiotic action was observed against Staphylococcus aureus and Escherichia coli. DVL also reduced antibiotic tolerance in S. aureus during 10 days of continuous treatment. In addition, DVL presented a nephroprotective effect, reducing sodium excretion, N-Gal expression and urinary protein, that are important markers of glomerular and tubular injuries. Taken together, studies of inhibition of hemagglutinating activity, fluorescence spectroscopy and molecular docking revealed that gentamicin can interact with DVL via the carbohydrate recognition domain (CRD), suggesting that the results obtained in this study may be directly related to the interaction of DVL-gentamicin and with the ability of the lectin to interact with glycans present in the cells of the peritoneum.

Collagen-silica nanocomposites as dermal dressings preventing infection in vivo.

The controlled delivery of multiple drugs from biomaterials is a timely challenge. In particular the nanocomposite approach offers a unique opportunity to combine the scaffold-forming ability and biocompatibility of hydrogels with the versatile and tunable drug release properties of micro- or nano-carriers. Here, we show that collagen-silica nanocomposites allowing for the prolonged release of two topical antibiotics are promising medicated dressings to prevent infection in wounds. For this purpose, core-shell silica particles loaded with gentamicin sulfate and sodium rifamycin were combined with concentrated collagen type I hydrogels. A dense fibrillar network of collagen exhibiting its typical periodic banding pattern and a homogenous particle distribution were observed by scanning electron microscopy. Antibiotics release from nanocomposites allowed a sustained antibacterial effect against Staphylococcus aureus over 10 days in vitro. The acute dermal irritation test performed on albino rabbit skin showed no sign of severe inflammation. The antibacterial efficiency of nanocomposites was evaluated in vivo in a model of cutaneous infection, showing a 2 log steps decrease in bacterial population when loaded systems were used. In parallel, the histological examination indicated the absence of M1 inflammatory macrophages in the wound bed after treatment. Taken together, these results illustrate the potentialities of the nanocomposite approach to develop collagen-based biomaterials with controlled dual drug delivery to prevent infection and promote cutaneous wound repair.

In vitro antibacterial activity of bioactive glass S53P4 on multiresistant pathogens causing osteomyelitis and prosthetic joint infection.

BACKGROUND: Conventional local treatment for medullary osteomyelitis (OM) includes insertion of antibiotic-loaded polymethylmethacrylate (PMMA) cement. Nevertheless, PMMA may delivery irregular concentration of antibiotic to surrounding tissue. We aimed to compare the in vitro antibacterial activity of Bioactive Glass (BAG) S53P4, which is a compound showing local antibacterial activity, to that of antibiotic-loaded PMMA against multidrug resistant bacteria from OM isolates. METHODS: We studied convenience samples of multidrug resistant (MDR) microorganisms obtained from patients presenting OM and prosthetic joint infection (PJI). Mixtures containing tryptic soy broth (TSB) and inert glass beads (2 mm), BAG-S53P4 granules (0.5-0.8 mm and < 45 mm) and Gentamicin or Vancomycin-loaded PMMA beads were inoculated with methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus (MR-CoNS), Pseudomonas aeruginosa or Klebsiella pneumoniae isolates. Glass beads (2.0 mm) were used as a control. Antibacterial activity was evaluated by means of time-kill curve, through seeding the strains on blood agar plates, and subsequently performing colony counts after 24, 48, 72, 96, 120 and 168 h of incubation. Differences between groups were evaluated by means of two-way analysis of variance (ANOVA) and Bonferroni's t test. RESULTS: Inhibition of bacterial growth started soon after 48 h of incubation, reached zero CFU/ml between 120 and 168 h of incubation for both antibiotic-loaded PMMA and BAG S53P4 groups, in comparison with inert glass (p < 0.05). No difference regarding time-kill curves between antibiotic-loaded PMMA and BAG S53P4 was observed. CONCLUSIONS: BAG S53P4 presented antibacterial properties as much as antibiotic-loaded PMMA for MDR bacteria producing OM and PJI.

Microbicidal gentamicin-alginate hydrogels.

Sodium alginate (Alg) reacted with antibiotic gentamicin sulfate (GS) in an aqueous-phase condition mediated by carbodiimide chemistry, in the molar ratios Alg: GS of (1:0.5), (1:1) and (1:2). The Alg-GS conjugated derivatives were characterized by elemental analysis for nitrogen content, Fourier transform infrared spectroscopy in the attenuated total reflection mode (FTIR-ATR), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), thermogravimetric analyses (TGA) and water sorption measurements. XPS and FTIR-ATR analyses clearly indicated that GS molecules covalently attached to the backbone of the alginate chains by amide bond formation. The highest amount of GS bound to Alg (43.5 ±â€¯0.4 wt%) and the highest swelling ratio (4962 ±â€¯661%) were observed for the Alg-GS (1:2) sample. Bioluminescence assays with Pseudomonas aeruginosa PAO1/lecA:lux and colony forming counting of Staphylococcus aureus and Escherichia coli upon contact with all Alg-GS conjugates revealed microbicidal activity; however, Alg-GS (1:2) was the most efficient, due to the highest GS content.

Spherical gold nanoparticles and gold nanorods for the determination of gentamicin.

Gentamicin is an antibiotic indicated to treat mastitis in dairy cattle and for the treatment of bacterial resistance in the context of hospital infections. The effect caused by gentamicin on the optical properties of gold nanoparticles aqueous dispersions were used to develop quantitative methods to determine this antibiotic. Two different aqueous dispersions, one containing spherical Au nanoparticles (AuNPs) and the other containing Au nanorods (AuNRs), had their conditions adjusted to enable a stable and sensitive response towards gentamicin. The use of AuNPs, with measurement at 681nm of the rising coupling plasmon band, enabled a limit of detection (LOD) of 0.4ngmL-1 (0.02ng absolute LOD), ten times lower than the one achieved by measuring the decreasing of the longitudinal surface plasmon resonance band (at 662nm). The linear analytical response of AuNPs measured at 681nm did not require rationing of signal values to correct for linearity. Stability of the analytical response resulted in intermediary precision below 2%. No significant interference was imposed by excipients traditionally present in injectable solutions for veterinary use. Percent recoveries obtained in such formulations were between 94.5 and 98.2% regardless the existence of any difference in the proportion of the compounds known as gentamicin (C1, C1a and C2) in standard and in the samples. The method requires no derivatization with toxic reagents as usually is required in other spectroscopic approaches.

Effects of the sol-gel route on the structural characteristics and antibacterial activity of silica-encapsulated gentamicin.

The effects of sol-gel processes, i.e., acid-catalyzed gelation, base-catalyzed gelation and base-catalyzed precipitation routes, on the encapsulation of gentamicin were investigated. The resulting xerogels were characterized using a series of complementary instrumental techniques, i.e., the adsorption/desorption of nitrogen, small-angle X-ray scattering, Fourier transform infrared spectroscopy, diffuse reflectance spectroscopy, X-ray photoelectron spectroscopy, atomic force microscopy and scanning electron microscopy. The encapsulated gentamicin samples were tested against a series of Gram-positive and Gram-negative bacterial strains. The best antimicrobial activity was observed with the encapsulated gentamicin that was prepared via the precipitation route, even in comparison with the neat antibiotic, especially in the case of the Gram-positive strain Staphylococcus aureus. The gentamicin concentration on the outermost surface and the zeta potential were identified as factors that affected the highest efficiency, as observed in the case of encapsulation via the base-catalyzed process.

Micro/nanostructured hyaluronic acid matrices with tuned swelling and drug release properties.

Hyaluronic acid (HA) hydrogels were structured in the form of porous monoliths by means of the ice-segregation-induced self-assembly (ISISA) method coupled with freeze-drying. Physical and chemical parameters were explored in order to fine-tune the microstructure and the incidence on both swelling and dissolution behavior in aqueous media. Gentamicin-loaded HA matrices with tuned drug release properties were also prepared; their inherent properties and behavior in solution are discussed in the framework of thermal analysis and scanning electron microscopy inspection.

Hydroxyapatite coating by sol-gel on Ti-6Al-4V alloy as drug carrier.

In this study Ti-6Al-4V samples were used as substrates and Ca-P layers were deposited using sol-gel technique and covered by spin-coating. The efficiency of hydroxyapatite (HA) coatings as drug carrier was also evaluated by immersion in gentamicin sulphate solution and the release profiles were obtained by cumulative method of the coating samples. Three non-linear mathematical methods were employed in order to discuss a possible mechanism to lead the drug release. Physical chemical techniques showed the presence of the typical absorption bands of calcium phosphates by infrared spectroscopy while X-ray diffraction peaks matched up with hydroxyapatite patterns. Microstructural techniques (SEM, EDS) help to confirm the hydroxyapatite coating by surface aspect and Ca/P ratio (1.64). The best fitting according statistical results explained each stage of the released profiles and correspond to a mixture of short initial burst effect plus drug dissolution with a specific kinetic and the diffusion of the gentamicin solid particles.

Analysis of underivatized gentamicin by capillary electrophoresis with UV detection.

A selective and rapid capillary zone electrophoresis method for determination of the multicomponent aminoglycoside antibiotic gentamicin is described. Baseline separation of gentamicin C1, C1a, C2, C2a and C2b components was achieved with a background electrolyte containing 0.35 mM cetyl trimethylammonium bromide, 3% methanol and 90 mM sodium pyrophosphate (pH 7.4) and detected directly with UV detection without derivatization. Quantitative analysis was performed and illustrated the potential use of capillary electrophoresis for the identification and quantitation of gentamicin components, but the application of this method is limited to a gentamicin concentration range of 2-6 mg/ml.

Nonstoichiometric hydroxyapatite-anionic collagen composite as support for the double sustained release of gentamicin and norfloxacin/ciprofloxacin.

This work studied the sustained release of ciprofloxacin or norfloxacin and gentamicin from nonstoichiometric hydroxyapatite (nHA) and anionic collagen composite. Within the first 24 and 48 h, the total antibiotic supply was significantly higher than the minimal inhibitory concentration required for the majority of the gram-negative bacteria. Although gentamicin was completely released from the matrix after 48 h by a normal diffusion mechanism, ciprofloxacin or norfloxacin release was characterized by a 2-phase release mechanism due to binding to nHA by complexation with calcium ion. Under the conditions studied, most of the norfloxacin or ciprofloxacin only will be disposable due to bioresorption or dissociation of the complexes. In conclusion, due to its biocompatibility nHA-anionic collagen composite may be a convenient support for the double sustained release of the antibiotics gentamicin and ciprofloxacin/norfloxacin for the control of bone infection while promoting bone tissue growth.

X-ray studies on phospholipid bilayers. XIII. Interactions with gentamicin.

This study deals with the structural perturbations that the aminoglycoside antibiotic gentamicin (GENT) can produce to phospholipid bilayers. Two multi-bilayer systems, one built-up of dimyristoylphosphatidylcholine (DMPC) and the other of dimyristoylphosphatidylethanolamine (DMPE) were allowed to interact with GENT. The experiments were performed in both a hydrophobic and a hydrophilic medium below the phospholipid main transition temperatures. X-ray diffraction techniques were used to determine the extent of the perturbation induced by GENT. The maximum effect was attained when GENT interacted with DMPC in the hydrophobic medium. On the other hand, GENT in aqueous solutions was unable to perturb in any significant extent the structure of the phospholipids under study.