In the past decades, titanium-based biomaterials have been broadly used in maxillofacial and periodontology surgery. The main aetiological agents related to complications in this procedures are Porphyromonas gingivalis, a Gram-negative anaerobic bacteria that is also responsible for the development of chronic gingivitis, and Streptococcus oralis, a Gram positive facultative anaerobic bacteria. In previous studies, we have demonstrated that the fluorine doping of titanium-based alloys reduces bacterial adherence.

The aim of this study is to evaluate the bacterial adherence on fluorine-doped titanium (TiF) probes compared to chemical polishing titanium (Ti) probes.

The P. gingivalis ATCC 33277 and S. oralis ATCC 9811 adherence study was performed by introducing each probe in a well of 6-well plate with 5 ml containing 106 colony forming units (CFU/ml) in sterile 0.9% NaCl and was incubated 37°C 5% CO2 for 90 minutes, in anaerobiosis in the case of P. gingivalis. After incubation, samples were stained with LIVE/DEAD BacLight Bacterial Viability Kit. Proportion of live and dead bacteria was calculated and studied by using ImageJ software. The experiments were performed in triplicate.

The statistical data were analyzed by nonparametric Wilcoxon test with a level of statistical significance of 0.05. Our results showed a significant (p<0.0053) 14.41% decrease of the adherence of P. gingivalis on TiF and an increase of 30% of dead cells. For S. oralis we did not get significant results.

In conclusion, TiF can be considered a promising approach to prevent and treat infections related to maxillofacial and periodontology surgery.

Prosthetic joint infections (PJI) occur infrequently, but they represent the most devastating complication with high morbidity and substantial cost. Staphylococcus aureus and coagulase-negative S. epidermidis are the most common infecting agents associated with PJI.

During the past decades, novel materials have been developed to improve osseointegration of implants. Recently has been demonstrated that by using nanosized hydroxyapatite (HA) coatings, since it combines nanoroughness and bone-like chemistry in a synergistic effect, it promotes better osseointegration when compared to uncoated metal implants. In a further step, due to the known bactericidal properties of fluor, the aim of this study is to evaluate the biofilm development on fluorohydroxyapatite (FHA) compared to HA.

Coatings were grown on stainless steel substrates by Pulsed Laser Deposition (PLD) technique using fluorohydroxyapatite targets of marine origin. A comprehensive physicochemical characterization of the coatings was performed using SEM, EDS, XPS and XRD. Biological in vitro tests using the pre-osteoblastic cell line (MC3T3-E1) demonstrated the non-cytotoxicity of FHA coatings, the healthy cell proliferation and their osteogenic activity.

The S. aureus 15981 (Valle et al.) and S. epidermidis ATCC 35984 strains adherence study was performed introducing each probe in a well of 96-well plate with 200 µl containing 106 colony forming units (CFU/mL) intryptic soy broth supplemented with 1% glucose and was incubated at 37°C 5% CO2 for 24 hours. After incubation, the medium was removed and three washes with 0.9% NaCl sterile saline were performed. The biofilm was disrupted by sonication at 50–60 Hz for 5 min. The CFU/cm2 was estimated by drop plate method. All of the experiments were performed in triplicate.

Statistical analysis was performed by non-parametric unilateral Wilcoxon”s test with a level of statistical significance of 0.05.

The results showed a significant (p=0.02475) 2.4-fold reduction in S. epidermidis biofilm formation on FHA (logUFC/cm2 = 6.87) compared to HA (logUFC/cm2 = 7.25); and also a significant (p=0.042) 3.7-fold reduction in S. aureus 15981 biofilm formation.

In conclusion, according our results FHA is a promising biomaterial that promotes osseointegration and decreases the staphylococcal biofilm that could avoid PJI. Further studies will be necessary.

Prosthetic joint infections (PJI) occur infrequently, but they represent the most devastating complication with high morbidity and substantial cost. Staphylococcus aureus and coagulase-negative S. epidermidis are the most commonly infecting agents associated with PJI. Nowadays, Gram-negative species like Escherichia coli and Pseudomonas aeruginosa are gaining relevance.

The use of TiO2 conical nanotubular doped with fluorine and phosphorous (FP-cNT) surfaces is an interesting approach to prevent surface bacterial colonization during surgery and favouring the osseointegration. Despite of there are serum markers related with PJI, to date there is described no biomaterial-related marker that allows detecting PJI. Here we describe the adherence and the bactericidal effect of FP-cNT and its capacity of marking the non-fermenting bacteria that have been in contact with it by Al. This metal is delivered by FP-cNT in non-toxic concentrations (between 25 and 29 ng/mL).

F-P-cNT layers on Ti6Al4V alloy were produced as described previously by our group. Ti6Al4V chemical polishing (CP) samples without nanostructure were used as control and produced as described previously.

S. aureus 15981, S. epidermidis ATCC 35984, E. coli ATCC 25922, and P. aeruginosa ATCC 27853 strains adherence study was performed using the protocol described by Kinnari et al. in 0.9% NaCl sterile saline with a 120 min incubation. After incubation, the samples were stained with LIVE/DEAD BacLight Bacterial Viability Kit. Proportion of live and dead bacteria was calculated and studied by using ImageJ software. The experiments were performed in triplicate. The aluminum concentration was estimated in the supernatant after incubation and in the 0.22 µm filtered supernatant by atomic absorption in graphite furnace.

The statistical data were analyzed by nonparametric Kruskal-Walis test and by pairwise comparisons using the nonparametric unilateral Wilcoxon test with a level of statistical significance of p<0.05. The values are cited as medians.

Our results show that the bacterial adherence of all tested species significantly decreased on FP-cNT compared to CP except P. aeruginosa ATCC 27853: 19.8% for S. aureus 15981, 45.3% for S. epidermidis ATCC 35984 and 8.1% for E. coli ATCC 25922. The bacterial viability decreased 2-fold for S. aureus 15981, and 5-fold for S. epidemidis ATCC 35984, but increased 95% for P. aeruginosa ATCC 27853 and there no was variation for E. coli ATCC 25922 on FP-cNT compared to CP. Only supernatant P. aeruginosa ATCC 27853 shows significant Al detection after 120 min incubation (p<0.05).

In summary, F-P cNT is a promising biomaterial that besides favoring osseointegration and potential usefulness as drug carrier, present bactericidal, non-stick ability (at least for staphylococci and E. coli) and is able to mark P. aeruginosa with Al, which could be potentially monitored in serum and urine in patients with PJI.

Background

Staphylococcus aureus is a human pathogen involved in implant-related infections. In these diseases, biofilm production is the key pathogenic event, and it increases antibiotic resistance of the organism. Because this phenomenon, local delivery of antibiotics could allows reaching high concentrations in the infected tissue without the secondary effects linked to systemic administration. Here we report the use of a ceramic biomaterial (SBA-15) as a carrier of antibiotics in order to deliver them directly in the infected tissue.

Objectives

Implant-related infection is one of the most devastating complications in orthopaedic surgery. Many surface and/or material modifications have been developed in order to minimise this problem; however, most of the in vitro studies did not evaluate bacterial adhesion in the presence of eukaryotic cells, as stated by the ‘race for the surface’ theory. Moreover, the adherence of numerous clinical strains with different initial concentrations has not been studied.

Introduction

Prosthetic joint infections (PJI) occur infrequently, but due to its increased clinical use represent the most devastating complication with high morbidity and substantial cost. Staphylococcus aureus and coagulase-negative staphylococci are the most common infecting agents associated with PJI. A possible therapeutic approach could be the local antibiotic by fluoride-TiO2 nanostructured anodic layers in order to prevent surface colonisation during the early moments after surgery. Here we describe the first results of this model using two common antibiotics.

Prosthetic joint infections (PJI) occur infrequently, but they represent the most devastating complication with high morbidity and substantial cost. Staphylococcus aureus are the most common infecting agents associated with acute PJI, and also appear in some cases of delayed PJI¹. S. aureus biofilm development can be divided in two stages: adhesion and proliferation². To avoid PJI bacterial adhesion has to be decreased.

Hybrid organo-inorganic sol-gel coatings are proposed as a promising biomaterial improvement³. One of these compounds is a mixture of two organopolisiloxanes: 3-methacryloxypropyltrimethoxysilane (MAPTMS) and tetramethylorthosilicate (TMOS). The aim of this work was to evaluate bacterial adhesion on MAPTMS-TMOS coating compared to titanium parts made by powder metallurgy.

MAPTMS-TMOS sol-gel coating was produced using a molar ratio of 1:2 (MAPTMS:TMOS) and dispersed in ethanol. The sol-gel was deposited by dip-coating on titanium parts made by powder metallurgy followed by a thermal treatment at 120 ºC for 30 minutes⁴. Titanium parts without sol-gel coating were used as control.

S. aureus 15981 strain adherence study was performed using the protocol described by Kinnari et al.⁵ with 90 min incubation. After incubation, the samples were stained with LIVE/DEAD BacLight Bacterial Viability Kit. Proportion of total adhered, live and dead bacteria was calculated and studied by using ImageJ software. The experiments were performed in triplicate.

The statistical data were analyzed by pairwise comparisons using the nonparametric Mann-Whitney test with a level of statistical significance of p<0.05. Values are cited and represented as medians.

S. aureus 15981 adherence was 942-fold lower on MAPTMS-TMOS coating than on uncoated titanium.

According with our results, MAPTMS-TMOS sol-gel coating is a promising antiadherent surface for S. aureus. More studies are necessary in order to evaluate this property with other species and strains.

Bone-regenerative and biocompatible materials are indicated for the regeneration of bone lost in periodontology and maxillofacial surgery. Bio-Oss is a natural bone mineral for bone grafting of bovine origin and the most common used in this kind of interventions¹. Sil-Oss is a new synthetic nanostructured monetite-based material that is reabsorbed at the same time that is replaced by new bone tissue ². Bacterial infection is one of the complications related to this kind of material. Streptococcus oralis is the most associated oral infecting pathogen to oral surgery³ and Staphylococcus aureus is the most common infecting pathogen to maxillofacial non-oral interventions⁴. Here we evaluated bacterial adherence of two of the most common infecting bacteria of this kind of biomaterial: S. oralis and S. aureus, on Bio-Oss and Sil-Oss.

S. oralis ATCC 9811 and S. aureus 15981 strains were used. Bacterial adherence was evaluated using the modified previously described protocol of Kinnari et al.⁵ that was adapted to our biomaterial. The quantification was performed by the drop plate method⁶. The statistical data were analyzed by pairwise comparisons using the nonparametric Mann-Whitney test with a level of statistical significance of p<0.05. Values are cited and represented as medians.

Bacterial adherence decreased significantly on Sil-Oss compared to Bio-Oss. S. oralis ATCC 9811 adherence was between 11 and 13-fold less on Sil-Oss compared to Bio-oss. In the case of S. aureus15981, the adherence was between 4 and 6-fold less on Sil-Oss compared to Bio-Oss.

Sil-Oss nanostructured monetite-based biomaterial could be considered as a promising biomaterial to be used for the regeneration of bone defects since the bacterial adherence on it is lower than on another currently used material.

Summary Statement

Description of an original in vitro protocol for assessing combined bacteria and cell competitive adherence on the surface of biomaterials of medical interest

Summary Statement

Combination of antibiotics with N-acetylcisteine and sub-MIC concentration of erythromycin was evaluated in two collection and 16 clinical strains of staphylococci isolated from PJI. The results were strain-dependent, so it evidences the necessity of perform individual studies of biofilm susceptibility.

Summary Statement

A study to evaluate biofilm development on different coatings of UHMWPE was performed. We observed a species-specific effect, with S. aureus affected mainly by DLC-F and S. epidermidis by DLC. These data correlates with previous adherence studies.

We have designed a prospective study to evaluate the usefulness of prolonged incubation of cultures from sonicated orthopaedic implants. During the study period 124 implants from 113 patients were processed (22 osteosynthetic implants, 46 hip prostheses, 54 knee prostheses, and two shoulder prostheses). Of these, 70 patients had clinical infection; 32 had received antibiotics at least seven days before removal of the implant. A total of 54 patients had sonicated samples that produced positive cultures (including four patients without infection). All of them were positive in the first seven days of incubation. No differences were found regarding previous antibiotic treatment when analysing colony counts or days of incubation in the case of a positive result. In our experience, extending incubation of the samples to 14 days does not add more positive results for sonicated orthopaedic implants (hip and knee prosthesis and osteosynthesis implants) compared with a conventional seven-day incubation period.

Cite this article: Bone Joint J 2013;95-B:1001–6.