Biofilms in Periprosthetic Orthopedic Infections Seen through the Eyes of Neutrophils: How Can We Help Neutrophils?

Despite advancements in our knowledge of neutrophil responses to planktonic bacteria during acute inflammation, much remains to be elucidated on how neutrophils deal with bacterial biofilms in implant infections. Further complexity transpires from the emerging findings on the role that biomaterials play in conditioning bacterial adhesion, the variety of biofilm matrices, and the insidious measures that biofilm bacteria devise against neutrophils. Thus, grasping the entirety of neutrophil-biofilm interactions occurring in periprosthetic tissues is a difficult goal. The bactericidal weapons of neutrophils consist of the following: ready-to-use antibacterial proteins and enzymes stored in granules; NADPH oxidase-derived reactive oxygen species (ROS); and net-like structures of DNA, histones, and granule proteins, which neutrophils extrude to extracellularly trap pathogens (the so-called NETs: an allusive acronym for "neutrophil extracellular traps"). Neutrophils are bactericidal (and therefore defensive) cells endowed with a rich offensive armamentarium through which, if frustrated in their attempts to engulf and phagocytose biofilms, they can trigger the destruction of periprosthetic bone. This study speculates on how neutrophils interact with biofilms in the dramatic scenario of implant infections, also considering the implications of this interaction in view of the design of new therapeutic strategies and functionalized biomaterials, to help neutrophils in their arduous task of managing biofilms.

Antibacterial Activity on Orthopedic Clinical Isolates and Cytotoxicity of the Antimicrobial Peptide Dadapin-1.

In orthopedic surgery, biomaterial-associated infections represent a complication of serious concern. Most promising strategies to prevent these infections currently rely on the use of anti-infective biomaterials. Desirably, in anti-infective biomaterials, the antibacterial properties should be achieved by doping, grafting, or coating the material surfaces with molecules that are alternative to conventional antibiotics and exhibit a potent and highly specific activity against bacteria, without altering the biocompatibility. Antimicrobial peptides (AMPs) are among the most interesting candidate molecules for this biomaterial functionalization. Here, the potential expressed by the recently discovered peptide Dadapin-1 was explored by assaying its MIC, MBIC and MBC on clinical strains of relevant bacterial species isolated from orthopedic infections and by assessing its cytotoxicity on the human osteoblast-like MG63 cells. When appropriately tested in diluted Mueller Hinton Broth II (MHB II), Dadapin-1 exhibited significant antibacterial properties. MIC values were in the range of 3.1-6.2 µM for the gram-positive bacteria Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus warneri, and 12.4-24.9 µM for the gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa. Interestingly, the peptide was found non-cytotoxic, with an IC50 exceeding the highest concentration tested of 179 µM. Overall, Dadapin-1 expresses considerable potential for future application in the production of anti-infective biomaterials.

Colonization and Infection of Indwelling Medical Devices by Staphylococcus aureus with an Emphasis on Orthopedic Implants.

The use of indwelling medical devices has constantly increased in recent years and has revolutionized the quality of life of patients affected by different diseases. However, despite the improvement of hygiene conditions in hospitals, implant-associated infections remain a common and serious complication in prosthetic surgery, mainly in the orthopedic field, where infection often leads to implant failure. Staphylococcus aureus is the most common cause of biomaterial-centered infection. Upon binding to the medical devices, these bacteria proliferate and develop dense communities encased in a protective matrix called biofilm. Biofilm formation has been proposed as occurring in several stages-(1) attachment; (2) proliferation; (3) dispersal-and involves a variety of host and staphylococcal proteinaceous and non-proteinaceous factors. Moreover, biofilm formation is strictly regulated by several control systems. Biofilms enable staphylococci to avoid antimicrobial activity and host immune response and are a source of persistent bacteremia as well as of localized tissue destruction. While considerable information is available on staphylococcal biofilm formation on medical implants and important results have been achieved on the treatment of biofilms, preclinical and clinical applications need to be further investigated. Thus, the purpose of this review is to gather current studies about the mechanism of infection of indwelling medical devices by S. aureus with a special focus on the biochemical factors involved in biofilm formation and regulation. We also provide a summary of the current therapeutic strategies to combat biomaterial-associated infections and highlight the need to further explore biofilm physiology and conduct research for innovative anti-biofilm approaches.

Different Sources of Mesenchymal Stem Cells for Tissue Regeneration: A Guide to Identifying the Most Favorable One in Orthopedics and Dentistry Applications.

The success of regenerative medicine in various clinical applications depends on the appropriate selection of the source of mesenchymal stem cells (MSCs). Indeed, the source conditions, the quality and quantity of MSCs, have an influence on the growth factors, cytokines, extracellular vesicles, and secrete bioactive factors of the regenerative milieu, thus influencing the clinical result. Thus, optimal source selection should harmonize this complex setting and ensure a well-personalized and effective treatment. Mesenchymal stem cells (MSCs) can be obtained from several sources, including bone marrow and adipose tissue, already used in orthopedic regenerative applications. In this sense, for bone, dental, and oral injuries, MSCs could provide an innovative and effective therapy. The present review aims to compare the properties (proliferation, migration, clonogenicity, angiogenic capacity, differentiation potential, and secretome) of MSCs derived from bone marrow, adipose tissue, and dental tissue to enable clinicians to select the best source of MSCs for their clinical application in bone and oral tissue regeneration to delineate new translational perspectives. A review of the literature was conducted using the search engines Web of Science, Pubmed, Scopus, and Google Scholar. An analysis of different publications showed that all sources compared (bone marrow mesenchymal stem cells (BM-MSCs), adipose tissue mesenchymal stem cells (AT-MSCs), and dental tissue mesenchymal stem cells (DT-MSCs)) are good options to promote proper migration and angiogenesis, and they turn out to be useful for gingival, dental pulp, bone, and periodontal regeneration. In particular, DT-MSCs have better proliferation rates and AT and G-MSC sources showed higher clonogenicity. MSCs from bone marrow, widely used in orthopedic regenerative medicine, are preferable for their differentiation ability. Considering all the properties among sources, BM-MSCs, AT-MSCs, and DT-MSCs present as potential candidates for oral and dental regeneration.

Cytocompatibility and antibacterial properties of capping materials.

The aim of this study was to evaluate and compare the antimicrobial activity and cytocompatibility of six different pulp-capping materials: Dycal (Dentsply), Calcicur (Voco), Calcimol LC (Voco), TheraCal LC (Bisco), MTA Angelus (Angelus), and Biodentine (Septodont). To evaluate antimicrobial activity, materials were challenged in vitro with Streptococcus mutans, Streptococcus salivarius, and Streptococcus sanguis in the agar disc diffusion test. Cytocompatibility of the assayed materials towards rat MDPC-23 cells was evaluated at different times by both MTT and apoptosis assays. Results significantly differed among the different materials tested. Both bacterial growth inhibition halos and cytocompatibility performances were significantly different among materials with different composition. MTA-based products showed lower cytotoxicity and valuable antibacterial activity, different from calcium hydroxide-based materials, which exhibited not only higher antibacterial activity but also higher cytotoxicity.

The interaction of bacteria with engineered nanostructured polymeric materials: a review.

Bacterial infections are a leading cause of morbidity and mortality worldwide. In spite of great advances in biomaterials research and development, a significant proportion of medical devices undergo bacterial colonization and become the target of an implant-related infection. We present a review of the two major classes of antibacterial nanostructured materials: polymeric nanocomposites and surface-engineered materials. The paper describes antibacterial effects due to the induced material properties, along with the principles of bacterial adhesion and the biofilm formation process. Methods for antimicrobial modifications of polymers using a nanocomposite approach as well as surface modification procedures are surveyed and discussed, followed by a concise examination of techniques used in estimating bacteria/material interactions. Finally, we present an outline of future sceneries and perspectives on antibacterial applications of nanostructured materials to resist or counteract implant infections.

Ozone Gel in Chronic Periodontal Disease: A Randomized Clinical Trial on the Anti-Inflammatory Effects of Ozone Application.

The search for new topical treatments able to display not only antimicrobial properties but also a multiplicity of other beneficial effects while expressing safe cytocompatibility toward host tissues is being progressively developed. Antiseptics represent an aid to the gold standard nonsurgical treatment Scaling-and-Root-Planing (SRP) for periodontal disease. This split-mouth study aims to assess the efficacy of the ozonized gel GeliO3 (Bioemmei Srl, Vicenza, Italy) plus SRP (experimental treatment), with respect to SRP + chlorhexidine gel. Ten participants were treated with SRP + chlorhexidine gel (control sites) and with SRP + ozone gel (trial sites). After 1 (T1) and 3 months (T2) from baseline (T0), patients were revisited. At each time-point, the following indexes were assessed: probing pocket depth (PPD), clinical attachment level (CAL), gingival index (GI), plaque index (PI), and bleeding on probing (BoP). It has been assessed that the use of the ozonized gel in addition to SRP did not show significant differences if compared to conventional SRP + chlorhexidine. Chlorhexidine was found to be more effective than ozone in reducing CAL and GI at T2. Ozone deserves consideration for its wide applicability in several clinical fields. In this connection, we also glance at the latest research on ozone therapy.

Copper-Alloy Surfaces and Cleaning Regimens against the Spread of SARS-CoV-2 in Dentistry and Orthopedics. From Fomites to Anti-Infective Nanocoatings.

The latest diffusion of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), responsible for the coronavirus disease (COVID-19), has involved the whole world population. Even if huge efforts to control the pandemic have been done, the viral spread is still continuing. COVID-19 is reported as a zoonosis jumped from bats and pangolins to humans. After infection in humans, SARS-CoV-2 is found in the nasopharyngeal and salivary secretions. The virus has also been detected in the blood plasma of infected patients. The viral spread occurs through droplets exhaled from the nose and mouth of the infected people when they breath or talk, or through droplets propelled as a dense cloud by chough or sneeze. The virus can also be delivered as an aerosol from blood plasma, through surgical procedures. Following these ways, the virus can disperse in the air, then reaching and settling on the exposed surfaces. How long the virus will survive on a surface depends on the material the surface is made from. Infection via high-touch surfaces should be prevented. Copper alloy coatings, combined with efficient hygienic/disinfectant procedures and careful surgical practice, could be helpful to health protection in dental practice and can also be adopted in orthopedic traumatology.

Ozonized Gel Against Four Candida Species: A Pilot Study and Clinical Perspectives.

Ozone therapy can display a wide range of clinical beneficial effects, including antimicrobial, immune-stimulant, analgesic, anti-hypoxic actions. However, there is still a paucity of data regarding the ozone fungicide activity. Oral Candida is the most common fungal infection in the mouth among denture wearers and people with weakened immune systems. In the case of generalized candidiasis or immunocompromised patients, systemic therapy is needed, while localized infections are treated with topic medications. However, many Candida strains are resistant to antifungal drugs. The aim of this preliminary analysis is to evaluate the antimycotic efficacy of a new ozonided oil (GeliO3), as a possible terapeutic alternative in local treatments of these infections, compared to chlorhexidine digluconate (Plak gel®). Chlorhexidine is a chemical synthesis disinfectant with a broad-spectrum antiseptic action, active against bacteria and fungi. Antimycotic activity was tested against the following four Candida species: C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, through an agar diffusion method. No significant differences were found between the growth inhibition zone diameters of the ozonized gel and chlorhexidine. The results indicated that the ozonized gel may help to combat Candida infections. Moreover, useful applications could be used to counteract Candida colonization of endosseous implants.

New concepts and new weapons in implant infections.

Molecular studies have cast light on new facts about the virulence factors of bacteria responsible for implant infections. Recently, the biofilm matrix has been shown to include a variety of important structural components, including DNA, in addition to polysaccharides and proteins. This finding has stimulated interest in substances able to disrupt biofilms by attacking both the poly-N-acetylglucosamine surface polysaccharide (PNAG) and the extracellular DNA (eDNA), namely, dispersin B, a PNAG-degrading enzyme, and DNase I. The therapeutic potential of these enzymes are reviewed in this issue of IJAO, as well as the ability of the excretions/secretions of the medicinal maggot Lucilia sericata to disrupt Staphylococcus epidermidis biofilms. The activity of bacterial proteases causes the release of eDNA, critical for the early development of biofilms. This complex process, including suicidal and fratricidal mechanisms, is also presented in the current issue of IJAO. The different sensitivities of S. aureus and S. epidermidis to enzymatic anti-biofilm agents and to the host's first line of defense, as well as the importance of knowing the cascade of regulatory genes in bacteria so as to interfere with biofilm production using gene therapy or quorum sensing inhibitors are also discussed in this issue. Other innovative approaches consist in disrupting biofilms by exposing them to photodynamic substances and simultaneously to visible light, and in coating biomaterial surfaces with organic molecules to prevent protein adsorption and biofilm formation.

Advancements in molecular epidemiology of implant infections and future perspectives.

Implant infection remains the major and often irreducible complication in clinical use of biomaterials, demanding new therapeutic and preventive strategies. Etio-pathogenesis of biomaterials-related infections is being more and more studied, and various virulence bacterial factors have progressively been identified, but little is still known about the weight of the distinct molecules in the context of specific peri-implant infection sites. Molecular epidemiology has become recently integrated into the research on implant infections. What distinguishes molecular epidemiology from the simple molecular biology is that the use of molecular techniques is applied to the study of the distribution and prevalence of virulence and resistance genes in collections of bacterial clinical isolates from implant infections. Here, the authors comment on the range of molecular techniques available, reviewing the various applications of molecular epidemiology to the study of implant infections and providing some experimental examples related to the field of orthopaedic implant infections. They highlight the new opportunities arising from molecular epidemiology of designing measures useful to prevent and treat implant infections. The knowledge of the relative weight of virulence factors and of their regulatory mechanisms at molecular level can open the way to new strategies also including gene therapies aimed at silencing or knocking out crucial genes responsible for the aggressive tools (adhesins, biofilm production, antibiotic resistance) of the aetiological agents of implant-related infections.

The significance of infection related to orthopedic devices and issues of antibiotic resistance.

Over the last 15 years, with the advent of modern standards in the control of sterility within the operating room environment and adequate protocols of peri-operative antibiotic prophylaxis, the incidence of infections associated to orthopedic implants has become very low. Nevertheless, the event of infection still represents one of the most serious and devastating complications which may involve prosthetic devices. It leads to complex revision procedures and, often, to the failure of the implant and the need for its complete removal. In orthopedics, for the enormous number of surgical procedures involving invasive implant materials, even if nowadays rare, infections have a huge impact in terms of morbidity, mortality, and medical costs. The difficult battle to prevent and fight bacterial infections associated to prosthetic materials must be played on different grounds. A winning strategy requires a clear view of the pathogenesis and the epidemiology of implant-related infections, with a special attention on the alarming phenomenon of antibiotic resistance. In this regard staphylococci are the prevalent and most important causative pathogens involved in orthopedic implant-related infections, and, thus, the main enemy to defeat. In this paper, we offer an overview of the complexity of this battleground and of the current and new, in our opinion most promising, strategies in the field of biomaterials to reduce the risks and counteract the establishment of implant infections.

Detection of biofilm formation in Staphylococcus epidermidis from implant infections. Comparison of a PCR-method that recognizes the presence of ica genes with two classic phenotypic methods.

Biofilm-forming ability is increasingly being recognized as an important virulence factor in Staphylococcus epidermidis. This study compares three different techniques for the detection of biofilm-positive strains. The presence of icaA and icaD genes responsible for biofilm synthesis was investigated by a PCR method in a collection of 80 S. epidermidis strains isolated from orthopedic implant infections. The results from molecular analysis were compared with those obtained by two classic phenotypic methods, the Congo red agar (CRA) plate test and the microtiter plate test (MtP). Fifty-seven percent of all the examined strains were found icaA/icaD-positive, of which only three were not positive for CRA test. Differently, by the MtP method, 66% of the strains were found to be biofilm-producers but only a limited agreement with the PCR-method was noticeable because of the observation of (icaA/icaD+)/MtP- strains (8%) and of a surprising ambiguous result of (icaA/icaD-)/MtP+ strains (16%). The category of the weak biofilm-producers provided the highest contribution to these mismatching results (10%). The better agreement between the CRA plate test with the molecular detection of ica genes indicates the former as a reliable test for the phenotypic characterization of virulence of clinical isolates. However, MtP method remains a precious tool for the in vitro screening of different biomaterials for the adhesive properties using a reference strain.

Effect of Winemaking on the Composition of Red Wine as a Source of Polyphenols for Anti-Infective Biomaterials.

Biomaterials releasing bactericides have currently become tools for thwarting medical device-associated infections. The ideal anti-infective biomaterial must counteract infection while safeguarding eukaryotic cell integrity. Red wine is a widely consumed beverage to which many biological properties are ascribed, including protective effects against oral infections and related bone (osteoarthritis, osteomyelitis, periprosthetic joint infections) and cardiovascular diseases. In this study, fifteen red wine samples derived from grapes native to the Oltrepò Pavese region (Italy), obtained from the winemaking processes of "Bonarda dell'Oltrepò Pavese" red wine, were analyzed alongside three samples obtained from marc pressing. Total polyphenol and monomeric anthocyanin contents were determined and metabolite profiling was conducted by means of a chromatographic analysis. Antibacterial activity of wine samples was evaluated against Streptococcus mutans, responsible for dental caries, Streptococcus salivarius, and Streptococcus pyogenes, two oral bacterial pathogens. Results highlighted the winemaking stages in which samples exhibit the highest content of polyphenols and the greatest antibacterial activity. Considering the global need for new weapons against bacterial infections and alternatives to conventional antibiotics, as well as the favorable bioactivities of polyphenols, results point to red wine as a source of antibacterial substances for developing new anti-infective biomaterials and coatings for biomedical devices.

Biofilm formation on titanium implants counteracted by grafting gallium and silver ions.

Biofilm-associated infections remain the leading cause of implant failure. Thanks to its established biocompatibility and biomechanical properties, titanium has become one of the most widely used materials for bone implants. Engineered surface modifications of titanium able to thwart biofilm formation while endowing a safe anchorage to eukaryotic cells are being progressively developed. Here surfaces of disks of commercial grade 2 titanium for bone implant were grafted with gallium and silver ions by anodic spark deposition. Scanning electron microscopy of the surface morphology and energy dispersive X-ray spectroscopy were used for characterization. Gallium-grafted titanium was evaluated in comparison with silver-grafted titanium for both in vivo and in vitro antibiofilm properties and for in vitro compatibility with human primary gingival fibroblasts. Surface-modified materials showed: (i) homogeneous porous morphology, with pores of micrometric size; (ii) absence of cytotoxic effects; (iii) ability to support in vitro the adhesion and spreading of gingival fibroblasts; and (iv) antibiofilm properties. Although both silver and gallium exhibited in vitro strong antibacterial properties, in vivo gallium was significantly more effective than silver in reducing number and viability of biofilm bacteria colonies. Gallium-based treatments represent promising titanium antibiofilm coatings to develop new bone implantable devices for oral, maxillofacial, and orthopedic applications.

Bacterial adhesion to poly-(D,L)lactic acid blended with vitamin E: toward gentle anti-infective biomaterials.

Anti-infective properties of biomedical materials are often achieved by loading or coating them with powerful bactericides. Undesirably, these bioactive molecules can damage the host cells at the biomaterial-tissues interface and, sometimes, even determine systemic toxic effects. The search for biomaterials able to actively resist infection while displaying a safe cytocompatibility profile toward eukaryotic cells is being progressively developed. Poly-(D,L)lactic acid (PLA) is a broadly used resorbable material with established biocompatibility properties. The dissolving surfaces of a biodegradable material tend to be per se elusive for bacteria. Here, films of pristine PLA, of PLA blended with vitamin E (VitE) and PLA blended with vitamin E acetate (VitE ac) were challenged in vitro with the biofilm-producers Staphylococcus epidermidis RP62A and Staphylococcus aureus ATCC25923. The bacterial adhesion properties of the different materials were investigated on small film disc specimens by a method based on microtiter plates. Adherent bacteria were quantified by both CFU plating and bioluminescence. Significant decrease in bacterial adhesion and biofilm accumulation was found on the surface of both the enriched polymers. These findings, together with the favorable intrinsic properties of PLA and the desirable bioactivities conferred by VitE, point up the VitE-blended PLA polymers as gentle anti-infective biomaterials.

Effects on antibiotic resistance of Staphylococcus epidermidis following adhesion to polymethylmethacrylate and to silicone surfaces.

A number of studies appears to give emphasis to the role of prosthetic materials in determining microbial adherence and resistance to host defence and drug therapy. Aim of this study was to explore whether the direct contact with biomaterial substrata of different chemical nature could influence bacterial behaviour, determining possible changes in the bacteria population as far as antibiotic resistance is concerned. To this end, susceptibility to penicillin, erythromycin, clindamycin, cefamandole, imipenem, vancomycin, ciprofloxacin. ampicillin, cefazolin, trimethoprim-sulfamethoxazole, chloramphenicol, amikacin and netilmicin was evaluated in a methicillin-, gentamicin- and tobramycin-resistant Staphylococcus epidermidis strain, after in vitro adhesion to polymethylmethacrylate (PM MA) and to silicone elastomer. The susceptibility to antibiotics of both adherent bacteria and bacteria which, although exposed to the materials, had not undergone adhesion was measured as bacterial growth inhibition area onto a plate antibiogram. according to Kirby-Bauer and using an image analyser system. The results obtained suggest that the two test materials considered in this study were capable to condition bacterial behaviour. In particular. the adhesion onto PMMA surfaces induced a marked and significant decrease in susceptibility to the following beta-lactam antibiotics: cefamandole (32%), cefazolin (23%), imipenem (27%), ampicillin (31%). Moreover, PMMA caused a lower but significant reduction in resistance to vancomycin (15%), chloramphenicol (16%), amikacin (13%). netilmicin (13%), erythromycin (11%) trimethoprim-sulfamethoxazole (13%). In contrast, the adhesion onto silicone elastomer appeared to influence bacterial changes to a lesser extent and elicited a significant decrease in susceptibility only to cefazolin (10%) and amikacin (11). Further studies are required to thoroughly investigate the mechanisms of these variations, even though, also according to other authors, one of the best conceivable conclusions is that some material substrata can lead to selection of variant adhesive bacteria with increased antibiotic resistance.

In vitro behaviour of bone marrow-derived mesenchymal cells cultured on fluorohydroxyapatite-coated substrata with different roughness.

Among ceramic materials, recent interest has been risen on fluorinated hydroxyapatites, which undergo slow in vivo degradation and offer a more stable interface for osseointegration and bone fixation. Apart from the information available on the chemistry, little is known of their biological properties and, more specifically, of their interaction with bone cells. The aim of the present study has been to investigate the behaviour and differentiation of human bone marrow-derived mesenchymal cells cultured on two substrata consisting of fluorohydroxyapatite (FHA)-coated titanium and characterised by different surface roughness. To this end, osteoprogenitor cells were seeded on the test materials and, once adhered, were induced to differentiate to osteoblastic cells. The cell behaviour on the different surfaces was monitored for a period of up to 2 weeks. The results obtained indicate that FHAs are cytocompatible materials, which allow the adhesion and growth of osteogenic cells. Mesenchymal cells promptly adhered, covering the surface of the test materials, and subsequently expressed some typical markers of osteoblasts. No significant difference in alkaline phosphatase specific activity was observed when comparing the two test material surfaces to plastic control. At 7 days the number of adhered cells and the presence in the medium of C-terminal propeptide of type I collagen appeared lower or slightly lower for cultures on FHA substrata than on plastic control. These differences tended to subside with time.

Search for the insertion element IS256 within the ica locus of Staphylococcus epidermidis clinical isolates collected from biomaterial-associated infections.

Staphylococcus epidermidis biofilm-forming strains produce a polysaccharide intercellular adhesin (PIA), which mediates bacterial cell aggregation and favours the colonisation on prosthetic implants. PIA synthesis is regulated by the icaADBC locus. In vitro, by repeated subcultures of a biofilm-producing strain, the loss of the ability to produce biofilm appears associated with the insertion of the IS256 element into the ica locus. This study was aimed (i) to investigate if the five genes of ica locus are always all present in different strains of S. epidermidis, and (ii) to search if IS256 insertion naturally occurs in ica locus without making recourse to the experimental procedure of repeated subcultures of strains. 120 S. epidermidis clinical isolates from peri-prosthesis infections were investigated both by an original multiplex PCR analysis of the ica genes and by PCR amplification of the IS256 element. Also two reference strains (the biofilm-negative S. epidermidis ATCC 12228 and the biofilm-forming ATCC 35984 [RP62A]) and two biofilm-negative RP62A-derived acriflavin mutants (D9 and HAM892) were analysed. D9 e HAM892 were for the first time shown to contain in ica locus, at the base 3319, a 1300-bp insertion with a DNA sequence corresponding to IS256. Among the 120 clinical isolates, 51 (43%) turned out completely ica-positive, 69 completely ica-negative (57%). The genes of the ica locus appear, in all cases of the present collection, strictly linked each other, so they are either all present or all absent. In this collection, IS256 was present in eight out of the 69 ica-negative strains and in 34 out of the 51 ica-positive strains. IS256, also when present in bacterial genomic DNA, was never found inside the ica locus, thus suggesting that insertion/excision of this element is not a natural occurring mechanism for off/on switching of biofilm production.

Evaluation of bacterial adhesion of Streptococcus mutans on dental restorative materials.

Bacterial adhesion to the surface of composite resins and other dental restorative materials is an important parameter in the aetiology of secondary caries formation. The aim of the present study was to investigate the adhesion of a Streptococcus mutans strain (ATCC 25175) on the surface of different restorative materials. The test materials examined included three flowable composites (Filtek Flow, Tetric Flow, and Arabesk Flow), three microhybrid composites (Clearfil APX, Solitaire 2, and Z250), two glass-ionomers (Fuji IX, Fuji IX fast), a compomer (F2000), an ormocer (Admira), and a control reference material (tissue culture grade, surface treated polystyrene). The adhesion tests were carried out in 24-well plates. Quantitative turbidimetric measurements were finally performed in order to indirectly evaluate the amount of bacteria retained on the material surface after in vitro exposure to the bacteria suspension. Under these conditions, with the exception of the Admira ormocer and the Fuji IX fast glass ionomer, which were found to be more adhesive, all the other material surfaces showed a similar susceptibility to bacterial adhesion, exhibiting values not significantly different than the reference polystyrene control. Furthermore, the release of fluoride from some of the test surfaces did not appear capable to reduce early bacterial adhesion.