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1.
Pathogens ; 12(2)2023 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-36839473

RESUMO

Arthroplasty is a highly successful treatment to restore the function of a joint. The contamination of the implant via bacterial adhesion is the first step toward the development of device-associated infections. The emerging concern about antimicrobial resistance resulted in a growing interest to develop alternative therapeutic strategies. Thus, the increment in the incidence of bacterial periprosthetic infections, the complexity of treating infections caused by organisms growing in biofilms, together with the rise in antibiotic resistant bacteria, expose the need to design novel surfaces that provide innovative solutions to these rising problems. The aim of this work is to develop a coating on titanium (Ti) suitable for inhibiting bacterial adhesion and proliferation, and hence, biofilm formation on the surface. We have successfully prepared polyacrylamide hydrogels containing the conventional antibiotic ampicillin (AMP), silver nanoparticles (AgNPs), and both, AMP and AgNPs. The release of the antibacterial agents from the gelled to aqueous media resulted in an excellent antibacterial action of the loaded hydrogels against sessile S. aureus. Moreover, a synergic effect was achieved with the incorporation of both AMP and AgNPs in the hydrogel, which highlights the importance of combining antimicrobial agents having different targets. The polyacrylamide hydrogel coating on the Ti surface was successfully achieved, as it was demonstrated by FTIR, contact angle, and AFM measurements. The modified Ti surfaces having the polyacrylamide hydrogel film containing AgNPs and AMP retained the highest antibacterial effect against S. aureus as it was found for the unsupported hydrogels. The modified surfaces exhibit an excellent cytocompatibility, since healthy, flattened MC3T3-E1 cells spread on the surfaces were observed. In addition, similar macrophage RAW 264.7 adhesion was found on all the surfaces, which could be related to a low macrophage activation. Our results indicate that AMP and AgNP-loaded polyacrylamide hydrogel films on Ti are a good alternative for designing efficient antibacterial surfaces having an excellent cytocompatibility without inducing an exacerbated immune response. The approach emerges as a superior alternative to the widely used direct adsorption of therapeutic agents on surfaces, since the antimicrobial-loaded hydrogel coatings open the possibility of modulating the concentration of the antimicrobial agents to enhance bacterial killing, and then, reducing the risk of infections in implantable materials.

2.
ACS Appl Bio Mater ; 4(8): 6451-6461, 2021 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-35006865

RESUMO

The use of implants in orthopedics and dental practice is a widespread surgical procedure to treat diverse diseases. However, peri-implantitis due to infections and/or poor osseointegration can lead to metallic implant failure. The aim of this study was to develop a multifunctional coating on titanium (Ti) surfaces, to simultaneously deal with both issues, by combining antibacterial silver nanoparticles (AgNPs) and regenerative properties of lactoferrin (Lf). A simple and cost-effective methodology that allows the direct multifunctionalization of Ti surfaces was developed. The modified surfaces were characterized by atomic force microscopy (AFM), X-ray photoelectron spectroscopy, and contact angle measurements. Additionally, in vitro preosteoblast cell adhesion, cell viability, and differentiation were evaluated. The antibacterial capability of the surfaces was tested against Staphylococcus aureus as a prosthesis infection model strain. Our results showed that Lf adsorbed on both Ti surfaces and Ti surfaces with adsorbed AgNPs. Simultaneously, the presence of Lf and AgNPs notably improved preosteoblast adhesion, proliferation, and differentiation, whereas it reduced the bacterial colonization by 97.7%. Our findings indicate that this simple method may have potential applications in medical devices to both improve osseointegration and reduce bacterial infection risk, enhancing successful implantation and patients' quality of life.


Assuntos
Nanopartículas Metálicas , Ortopedia , Antibacterianos/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Humanos , Nanopartículas Metálicas/química , Osseointegração , Próteses e Implantes , Qualidade de Vida , Prata/farmacologia , Titânio/farmacologia
3.
Colloids Surf B Biointerfaces ; 198: 111456, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33246776

RESUMO

The tuning of surface properties through functionalization is an important field of research with a broad spectrum of applications. Self-assembled monolayers (SAMs) allow the surface tailoring through the adsorption of molecular layers having the appropriate functional group or precursor group enabling in situ chemical reactions and thus to the incorporation of new functionalities. The latter approach is particularly advantageous when the incorporation of huge groups is needed. In this study, we report the immobilization of pterin moieties on 11-bromoundecyltrichlorosilane-modified silicon substrates based on the in situ replacement of the bromine groups by pterin (Ptr), the parent derivative of pterins, by means of a nucleophilic substitution reaction. The modified surface was structurally characterized through a multi-technique approach, including high-resolution XPS analysis, contact angle measurements, and AFM. The designed synthesis method leads to the functionalization of the silicon surface with two compounds, O-undecyl-Ptr and N-undecyl-Ptr, with a higher proportion of the N-derivative (1:8 ratio). The alkyl-pterins immobilized via the proposed strategy, retain their photochemical properties, being able to inhibit Staphylococcus aureus growth under irradiation (84.3 ± 15.6 % reduction in viable cells). Our results open the possibility for the modification of several materials, such as glass and metal, through the formation of SAMs having the proper head group, thus allowing the design of photosensitive surfaces with potential microbiological self-cleaning properties.


Assuntos
Silício , Staphylococcus aureus , Fármacos Fotossensibilizantes , Pterinas , Propriedades de Superfície
4.
ACS Biomater Sci Eng ; 5(10): 4920-4936, 2019 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-33455240

RESUMO

Device-associated infections (DAI) remain a serious concern in modern healthcare. Bacterial attachment to a surface is the first step in biofilm formation, which is one of the main causes of DAIs. The development of materials capable of preventing or inhibiting bacterial attachment constitutes a promising approach to deal with this problem. The multifactorial nature of biofilm maturation and antibiotic resistance directs the research for multitargeted or combinatorial therapeutic approaches. One attractive strategy is the modification or the engineering of surfaces in order to provide antiadhesive and/or antimicrobial properties. Currently, several different approaches that involve physical and chemical surface modification deliver some possible alternatives to achieve this goal. The engineered surfaces can be coated with molecules capable of inhibiting the bacterial adhesion or with active agents that kill microorganisms. In addition, surfaces can also be modified in order to be stimuli-responsive, responding to a particular trigger and then delivering the consequent antimicrobial outcome. Here, we review the prevailing strategies to modify surfaces in order to create an antimicrobial surface and discuss how different surface functionalization can affect bacterial adhesion and/or viability.

5.
ACS Appl Mater Interfaces ; 10(28): 23657-23666, 2018 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-29927235

RESUMO

The increasing incidence of infections in implantable devices has encouraged the search for biocompatible antimicrobial surfaces. To inhibit the bacterial adhesion and proliferation on biomaterials, several surface functionalization strategies have been developed. However, most of these strategies lead to bacteriostatic effect and only few of these are able to reach the bactericidal condition. In this work, bactericidal surfaces were designed through the functionalization of titanium surfaces with poly-l-lysine (PLL) as the mediator for the incorporation of antimicrobial silver nanoparticles (AgNPs). This functionalization influences the adsorption of the particles on the substrate impeding the agglomeration observed when bare titanium surfaces are used, leading to a homogeneous distribution of AgNPs on the surfaces. The antimicrobial activity of this surface has been tested against two different strains, namely, Staphylococcus aureus and Pseudomonas aeruginosa. For both strains and different AgNPs sizes, the surface modified with PLL and AgNPs shows a much enhanced antimicrobial activity in comparison with AgNPs deposited on bare titanium. This enhanced antibacterial activity is high enough to reach bactericidal effect, a condition hard to achieve in antimicrobial surfaces. Importantly, the designed surfaces are able to decrease the bacterial viability more than 5 orders with respect to the initial bacterial inoculum. That means that a relative low load of AgNPs on the PLL-modified titanium surfaces reaches 99.999% bacterial death after 24 h. The results of the present study are important to avoid infections in indwelling materials by reinforcing the preventive antibiotic therapy usually dosed throughout the surgical procedure and during the postoperative period.


Assuntos
Polilisina/química , Antibacterianos , Anti-Infecciosos , Nanopartículas Metálicas , Testes de Sensibilidade Microbiana , Prata
6.
Colloids Surf B Biointerfaces ; 164: 262-271, 2018 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-29413605

RESUMO

The use of indwelling devices has emerged as a frequent and often life-saving medical procedure. However, infection in prosthetic surgery is one of the most important and devastating complications. Once the biofilm has been formed, its eradication is extremely difficult, due to an increased resistance to host defense and conventional antimicrobials. Thus, the design of novel strategies for inhibiting the bacterial adhesion on implantable devices is a key point for successful surgical procedures. In this work, the development of a simple two-step protocol to prepare surfaces able to prevent the bacterial growth was successfully achieved. The surface-modification design includes a combined approach involving the multi-functionalization of Ti surfaces with silver nanoparticles (AgNPs) and/or ampicillin (AMP). The surface chemistry involved in AMP adsorption on titanium and silver surfaces was elucidated for the first time, thus establishing the basis for the further anchoring of other antibacterial compounds having similar functional groups. Our results show that the antibiotic binds to the titanium surface through covalent interactions between the COOH groups in AMP and the OH groups of the native TiO2 on the surface, although electrostatic interactions between protonated AMP and negatively charged TiO2 can also contribute to the antibiotic anchoring to the surface. The AMP immobilization on the AgNPs is carried out by thiolate-like bonds. The ß-lactam ring functionality is preserved after the adsorption process, since the Ti-AgNPs-AMP surface was able to decrease the bacterial viability in more than 80%. Moreover, the antimicrobial capacity is maintained over time due to a two-pathway antibacterial mechanism: death by contact (AMP) and death by release (AgNPs). The effect of AMP prevails on AgNPs at early stages of bacterial adhesion, while AgNPs are responsible for sustaining the relatively low but steady release of Ag(I), preserving the bacteriostatic activity of the surface over time. This effect would contribute to prevent infections due to sessile cells on indwelling devices, powering the action of the immune system and the conventional antibiotics usually dosed in implanted patients.


Assuntos
Anti-Infecciosos/farmacologia , Materiais Biocompatíveis/farmacologia , Bactérias Gram-Positivas/efeitos dos fármacos , Adsorção , Ampicilina/química , Ampicilina/farmacologia , Anti-Infecciosos/química , Ácido Cítrico/química , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Espectroscopia Fotoeletrônica , Prata/farmacologia , Espectrofotometria Ultravioleta , Titânio/farmacologia
7.
Nanoscale ; 7(38): 15789-97, 2015 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-26355753

RESUMO

The insertion and function of the muscle-type nicotinic acetylcholine receptor (nAChR) in Au(111)-supported thiolipid self-assembled monolayers have been studied by atomic force microscopy (AFM), surface plasmon resonance (SPR), and electrochemical techniques. It was possible for the first time to resolve the supramolecular arrangement of the protein spontaneously inserted in a thiolipid monolayer in an aqueous solution. Geometric supramolecular arrays of nAChRs were observed, most commonly in a triangular form compatible with three nAChR dimers of ∼20 nm each. Addition of the full agonist carbamoylcholine activated and opened the nAChR ion channel, as revealed by the increase in capacitance relative to that of the nAChR-thiolipid system under basal conditions. Thus, the self-assembled system appears to be a viable biomimetic model to measure ionic conductance mediated by ion-gated ion channels under different experimental conditions, with potential applications in biotechnology and pharmacology.


Assuntos
Proteínas de Peixes/química , Ouro/química , Receptores Nicotínicos/química , Compostos de Sulfidrila/química , Animais , Carbacol , Proteínas de Peixes/metabolismo , Microscopia de Força Atômica , Receptores Nicotínicos/metabolismo , Torpedo
8.
J Colloid Interface Sci ; 433: 86-93, 2014 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-25112916

RESUMO

The adsorption of flagellin monomers from Pseudomonas fluorescens on Au(111) has been studied by Atomic Force Microscopy (AFM), Scanning Tunneling Microscopy (STM), X-ray Photoelectron Spectroscopy (XPS), Surface Plasmon Resonance (SPR), and electrochemical techniques. Results show that flagellin monomers spontaneously self-assemble forming a monolayer thick protein film bounded to the Au surface by the more hydrophobic subunit and exposed to the environment the hydrophilic subunit. The films are conductive and allow allocation of electrochemically active cytochrome C. The self-assembled films could be used as biological platforms to build 3D complex molecular structures on planar metal surfaces and to functionalize metal nanoparticles.


Assuntos
Flagelina/química , Ouro/química , Membranas Artificiais , Nanopartículas Metálicas/química , Multimerização Proteica , Pseudomonas fluorescens/química , Ressonância de Plasmônio de Superfície/métodos
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