Dynamic Adhesive Behavior and Biofilm Formation of Staphylococcus aureus on Polylactic Acid Surfaces in Diabetic Environments.

Interest in biodegradable implants has focused attention on the resorbable polymer polylactic acid. However, the risk of these materials promoting infection, especially in patients with existing pathologies, needs to be monitored. The enrichment of a bacterial adhesion medium with compounds that are associated with human pathologies can help in understanding how these components affect the development of infectious processes. Specifically, this work evaluates the influence of glucose and ketone bodies (in a diabetic context) on the adhesion dynamics of S. aureus to the biomaterial polylactic acid, employing different approaches and discussing the results based on the physical properties of the bacterial surface and its metabolic activity. The combination of ketoacidosis and hyperglycemia (GK2) appears to be the worst scenario: this system promotes a state of continuous bacterial colonization over time, suppressing the stationary phase of adhesion and strengthening the attachment of bacteria to the surface. In addition, these supplements cause a significant increase in the metabolic activity of the bacteria. Compared to non-enriched media, biofilm formation doubles under ketoacidosis conditions, while in the planktonic state, it is glucose that triggers metabolic activity, which is practically suppressed when only ketone components are present. Both information must be complementary to understand what can happen in a real system, where planktonic bacteria are the ones that initially colonize a surface, and, subsequently, these attached bacteria end up forming a biofilm. This information highlights the need for good monitoring of diabetic patients, especially if they use an implanted device made of PLA.

Modification of physico-chemical surface properties and growth of Staphylococcus aureus under hyperglycemia and ketoacidosis conditions.

Diabetes is a widely spread disease affecting the quality of life of millions of people around the world and is associated to a higher risk of developing infections in different parts of the body. The reasons why diabetes enhances infection episodes are not entirely clear; in this study our aim was to explore the changes that one of the most frequently pathogenic bacteria undergoes when exposed to hyperglycemia and ketoacidosis conditions. Physical surface properties such as hydrophobicity and surface electrical charge are related to bacterial growth behavior and the ability of Staphylococcus aureus to form biofilms. The addition of glucose made bacteria more negatively charged and with moderate-intermediate hydrophobicity. Ketone bodies increased hydrophobicity to approximately 75% and pathological concentrations hindered some of the bacterial surface charge by decreasing the negative zeta potential of cells. When both components were present, the bacterial physical surface changes were more similar to those observed in ketone bodies, suggesting a preferential adsorption of ketone bodies over glucose because of the more favorable solubility of glucose in water. Glucose diabetic concentrations gave the highest number of bacteria in the stationary phase of growth and provoked an increase in the biofilm slime index of around 400% in relation to the control state. Also, this situation is related with an increase of bacterial coverage. The combination of a high concentration of glucose and ketone bodies, which corresponds to a poorly controlled diabetic situation, appears associated with an early infection phase; increased hydrophobic attractive force and reduced electrostatic repulsion between cells results in better packing of cells within the biofilm and more efficient retention to the host surface. Knowledge of bacterial response in high amount of glucose and ketoacidosis environments can serve as a basis for designing strategies to prevent bacterial adhesion, biofilm formation and, consequently, the development of infections.

The role of magnesium in biomaterials related infections.

Magnesium is currently increasing interest in the field of biomaterials. An extensive bibliography on this material in the last two decades arises from its potential for the development of biodegradable implants. In addition, many researches, motivated by this progress, have analyzed the performance of magnesium in both in vitro and in vivo assays with gram-positive and gram-negative bacteria in a very broad range of conditions. This review explores the extensive literature in recent years on magnesium in biomaterials-related infections, and discusses the mechanisms of the Mg action on bacteria, as well as the competition of Mg2+ and/or synergy with other divalent cations in this subject.

In vivo bactericidal efficacy of the Ti6Al4V surface after ultraviolet C treatment.

BACKGROUND: Biomaterial-associated infections are one of the most important complications in orthopedic surgery. The main goal of this study was to demonstrate the in vivo bactericidal effect of ultraviolet (UV) irradiation on Ti6Al4V surfaces. MATERIALS AND METHODS: An experimental model of device-related infections was developed by direct inoculation of Staphylococcus aureus into the canal of both femurs of 34 rats. A UV-irradiated Ti6Al4V pin was press-fit into the canal by retrograde insertion in one femur and the control pin was inserted into the contralateral femur. To assess the efficacy of UV radiation, the mean colony counts after inoculation in the experimental subjects and the control group were compared at different times of sacrifice and at different inoculum doses. RESULTS: At 72 h, the mean colony counts after inoculation in experimental femurs were significantly lower than those of the control group, with a reduction percentage of 76 % (p = 0.041). A similar difference between control and experimental pins was observed at 24 h using an inoculum dose <104 colony-forming units (CFU), for which the reduction percentage was 70.48 % (p = 0.017). CONCLUSION: The irradiated surface of Ti6Al4V is able to reduce early bacterial colonization of Ti6AlV pins located in the medullar channel and in the surrounding femur. The reductions depend on the initial inoculums used to cause infection in the animals and the greatest effects are detected for inoculums <104 CFU. LEVEL OF EVIDENCE: Not applicable.

Quercitrin-nanocoated titanium surfaces favour gingival cells against oral bacteria.

Many dental implants fail due to the infection and inflammation that walk hand in hand with poor healing and soft tissue integration. Titanium surfaces were nanocoated with quercitrin, a natural flavonoid, with the aim to improve soft tissue integration and increase dental implants success. Streptococcus mutans attachment and biofilm formation was analysed. Then, the anti-inflammatory properties and the potential of quercitrin-nanocoated surfaces to boost soft tissue regeneration were tested using human gingival fibroblasts. An inflammatory situation was mimicked using interleulin-1-beta. We found that quercitrin-nanocoated surfaces decreased initial bacterial adhesion while increasing human gingival fibroblasts attachment. Furthermore, quercitrin-nanocoated Ti increased collagen mRNA levels and decreased matrix metalloproteinase-1/tissue inhibitor of metalloproteinanse-1 mRNA ratio, which is related to a reduced metalloproteinase-mediated collagen degradation, while also decreasing the pro-inflammatory prostaglandin E2 release under basal and inflammatory conditions. These results suggest that quercitrin-nanocoated surfaces could enhance the soft tissue integration and increase dental implants success.

Electrochemical analysis of the UV treated bactericidal Ti6Al4V surfaces.

This research investigates in detail the bactericidal effect exhibited by the surface of the biomaterial Ti6Al4V after being subjected to UV-C light. It has been recently hypothesized that small surface currents, occurring as a consequence of the electron-hole pair recombination taking place after the excitation process, are behind the bactericidal properties displayed by this UV-treated material. To corroborate this hypothesis we have used different electrochemical techniques, such as electrochemical impedance spectroscopy (EIS), potentiodynamic polarization plots and Mott-Schottky plots. EIS and Mott-Schottky plots have shown that UV-C treatment causes an initial increase on the surface electrical conduction of this material. In addition, EIS and polarization plots demonstrated that higher corrosion currents occur at the UV treated than at the non-irradiated samples. Despite this increase in the corrosion currents, EIS has also shown that such currents are not likely to affect the good stability of this material oxide film since the irradiated samples completely recovered the control values after being stored in dark conditions for a period not longer than 24h. These results agree with the already-published in vitro transitory behavior of the bactericidal effect, which was shown to be present at initial times after the biomaterial implantation, a crucial moment to avoid a large number of biomaterial associated infections.

Adsorption behavior of human plasma fibronectin on hydrophobic and hydrophilic Ti6Al4V substrata and its influence on bacterial adhesion and detachment.

Biomaterial implant-associated infections, a common cause of medical devices' failure, are initiated by bacterial adhesion to an adsorbed protein layer on the implant material surface. In this study, the influence of protein surface orientation on bacterial adhesion has been examined using three clinically relevant bacterial strains known to express specific binding sites for human plasma fibronectin (HFN). HFN was allowed to adsorb on hydrophobic Ti6Al4V and physically modified hydrophilic Ti6Al4V substrata. Ellipsometric data reveal that the characteristics of the adsorbed protein layers primary depend on solid surface tension and the initial protein concentration in solution. In particular, HFN molecules adopt a more extended conformation on hydrophobic than hydrophilic surfaces, an effect that is more pronounced at low than at high initial protein concentrations. Moreover, the extended conformation of the protein molecules on these surfaces likely facilitates the exposure of specific sites for adhesion, resulting in the higher bacterial-cell attachment observed regardless of the strain considered. Contact angle measurements and the analysis of the number of remaining adhering cells after being subjected to external forces further suggest that both specific and nonspecific (hydrophobic) interactions play an important role on bacterial attachment. This study is the first one to evaluate the influence of surface hydrophobicity on protein adsorption and its subsequent effect on bacterial adhesion using a material whose hydrophobicity was not modified using chemical treatments that potentially led to surface properties changes other than hydrophobicity.

Bactericidal behaviour of Ti6Al4V surfaces after exposure to UV-C light.

TiO(2)-coated biomaterials that have been excited with UV irradiation have demonstrated biocidal properties in environmental applications, including drinking water decontamination. However, this procedure has not been successfully applied towards the killing of pathogens on medical titanium-based implants, mainly because of practical concerns related to irradiating the inserted biomaterial in situ. Previous researchers assumed that the photocatalysis on the TiO(2) surface during UV application causes the bactericidal effects. However, we show that a residual post-irradiation bactericidal effect exists on the surface of Ti6Al4V, not related with photocatalysis. Using a combination of staining, serial dilutions, and a biofilm assay, we show a significant and time-dependent loss in viability of different bacterial strains of Staphylococcus epidermidis and Staphylococcus aureus on the post-irradiated surface. Although the duration of this antimicrobial effect depends on the strains selected, our experiments suggest that the effect lasts at least 60 min after surface irradiation. The origin of such phenomena is discussed in terms of the physical properties of the irradiated surfaces, which include the emission of energy and changes in surfaces charge occurring during electron-hole recombination processes. The method here proposed for the preparation of antimicrobial titanium surfaces could become especially useful in total implant surgery for which the antimicrobial challenge is mainly during or shortly after surgery.

In vitro biocompatibility and bacterial adhesion of physico-chemically modified Ti6Al4V surface by means of UV irradiation.

UV irradiation leads to a "spontaneous" wettability increase of the Ti6Al4V surface while preserving bulk properties of the alloy that are crucial for its performance as an orthopedic and dental implant. We hypothesized that UV treatment of Ti6Al4V may impair bacterial adhesion without compromising the good response of human bone-forming cells to this alloy. The in vitro biocompatibility of the Ti6Al4V surface, before and after UV irradiation, was analyzed by using human cells related to the osteoblastic phenotype. The adhesion processes of bacterial strains related to clinical orthopedic infections, i.e., Staphylococcus aureus and Staphylococcus epidermidis, were studied theoretically and in vitro, under dynamic and static conditions as well as in the presence or absence of shear forces. While human cell adhesion was not altered by UV irradiation of Ti6Al4V alloy, this treatment reduced not only initial bacterial adhesion rates but also the number of bacteria retained on the surface after the passage of two air-liquid interfaces on the previously adhered bacteria. This study proposes the use of UV treatment prior to implantation protocols as an easy, economic and effective way of reducing bacterial adhesion on the Ti6Al4V surface without compromising its excellent biocompatibility.