Christian Vassena

Antimicrobial activity of pure platelet-rich plasma against microorganisms isolated from oral cavity

BackgroundAutologous platelet concentrates (PCs) have been extensively used in a variety of medical fields to promote soft and hard tissue regeneration. The significance behind their use lies in the abundance of growth factors in platelets α-granules that promotes wound healing. In addition, antibacterial properties of PCs against various bacteria have been recently pointed out. In this study, the antimicrobial effect of pure platelet-rich plasma (P-PRP) was evaluated against oral cavity microorganisms such as Enterococcus faecalis, Candida albicans, Streptococcus agalactiae, Streptococcus oralis and Pseudomonas aeruginosa. Blood samples were obtained from 17 patients who underwent oral surgery procedures involving the use of P-PRP. The antibacterial activity of P-PRP, evaluated as the minimum inhibitory concentration (MIC), was determined through the microdilution twofold serial method.ResultsP-PRP inhibited the growth of Enterococcus faecalis, Candida albicans, Streptococcus agalactiae and Streptococcus oralis, but not of Pseudomonas aeruginosa strains.ConclusionsP-PRP is a potentially useful substance in the fight against postoperative infections. This might represent a valuable property in adjunct to the enhancement of tissue regeneration.

A comparative study of the use of bioactive glass S53P4 and antibiotic-loaded calcium-based bone substitutes in the treatment of chronic osteomyelitis: a retrospective comparative study

The treatment of chronic osteomyelitis often includes surgical debridement and filling the resultant void with antibiotic-loaded polymethylmethacrylate cement, bone grafts or bone substitutes. Recently, the use of bioactive glass to treat bone defects in infections has been reported in a limited series of patients. However, no direct comparison between this biomaterial and antibiotic-loaded bone substitute has been performed. In this retrospective study, we compared the safety and efficacy of surgical debridement and local application of the bioactive glass S53P4 in a series of 27 patients affected by chronic osteomyelitis of the long bones (Group A) with two other series, treated respectively with an antibiotic-loaded hydroxyapatite and calcium sulphate compound (Group B; n = 27) or a mixture of tricalcium phosphate and an antibiotic-loaded demineralised bone matrix (Group C; n = 22). Systemic antibiotics were also used in all groups. After comparable periods of follow-up, the control of infection was similar in the three groups. In particular, 25 out of 27 (92.6%) patients of Group A, 24 out of 27 (88.9%) in Group B and 19 out of 22 (86.3%) in Group C showed no infection recurrence at means of 21.8 (12 to 36), 22.1 (12 to 36) and 21.5 (12 to 36) months follow-up, respectively, while Group A showed a reduced wound complication rate. Our results show that patients treated with a bioactive glass without local antibiotics achieved similar eradication of infection and less drainage than those treated with two different antibiotic-loaded calcium-based bone substitutes.

Use of Dithiothreitol to Improve the Diagnosis of Prosthetic Joint Infections

Diagnosis of prosthetic joint infections (PJIs) remains a challenge for microbiologists, despite new techniques for bacteria isolation have been developed in recent years. A widely recognized standard method has not yet been indicated mainly because of limitations due difficult procedures and need of dedicated instrumentation. We evaluated the ability of a sulfhydryl compound routinely used in microbiology laboratories, dithiothreitol (DTT), to dislodge bacteria from biofilm, keeping them alive and cultivable for identification and antibiotic susceptibility testing. We compared DTT treatment against sonication of prosthesis and culture of periprosthetic tissues, in order to establish if it could be introduced in routine microbiological diagnosis of PJIs. The study was conducted on 76 patients, 34 with aseptic loosening of their prosthesis and 42 who were diagnosed for PJI. DTT treatment gave results similar to sonication in terms of bacterial yielding. Sonication provided higher sensitivity (71.4%) and specificity (94.1%) respect to periprosthetic tissue culture, while DTT showed the same specificity of sonication but a better sensitivity (85.7%), especially when the causative microorganism was Staphylococcus epidermidis. In conclusion, we demonstrated that DTT could be used for PJIs diagnosis, thanks to its ease of use and its high sensitivity and specificity.

Photodynamic antibacterial and antibiofilm activity of RLP068/Cl against Staphylococcus aureus and Pseudomonas aeruginosa forming biofilms on prosthetic material

Prosthetic joint infections (PJIs) are becoming a growing public health concern in developed countries as more people undergo arthroplasty for bone fixation or joint replacement. Because a wide range of bacterial strains responsible for PJIs can produce biofilms on prosthetic implants and because the biofilm structure confers elevated bacterial resistance to antibiotic therapy, new drugs and therapies are needed to improve the clinical outcome of treatment of PJIs. Antimicrobial photodynamic therapy (APDT), a non-antibiotic broad-spectrum antimicrobial treatment, is also active against multidrug-resistant micro-organisms such as meticillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. APDT uses a photosensitiser that targets bacterial cells following exposure to visible light. APDT with RLP068/Cl, a novel photosensitiser, was studied by confocal laser scanning microscopy (CLSM) to evaluate the disruption of MRSA and P. aeruginosa biofilms on prosthetic material. Quantitative CLSM studies showed a reduction in biofilm biomass (biofilm disruption) and a decrease in viable cell numbers, as determined by standard plate counting, in the S. aureus and P. aeruginosa biofilms exposed to APDT with the photosensitiser RLP068/Cl. APDT with RLP068/Cl may be a useful approach to the treatment of PJI-associated biofilms.

In vitro antibiofilm activity of bioactive glass S53P4

AIM This work aimed to investigate the ability of different formulations of bioactive glass (BAG)-S53P4 to interfere with bacterial biofilm produced on prosthetic material by methicillin-resistant Staphylococcus aureus and multi-drug-resistant Pseudomonas aeruginosa. MATERIALS & METHODS Antibiofilm activity of three formulations of bioglass was assessed at different time points through two different analyses: Crystal Violet and confocal laser scanning microscopy assays. RESULTS Significant differences in the whole biofilm were observed between BAG-S53P4-treated and control samples, while no marked changes in antibiofilm activity were observed among the tested formulations. Data from colorimetric assay were confirmed by confocal laser scanning microscopy analysis, which evidenced the significant reduction in biomass and a decrease of total cell volume when both S. aureus and P. aeruginosa biofilms were treated with BAG-S53P4. CONCLUSION BAG-S53P4 can be considered as an excellent adjuvant in the treatment of prosthetic infections related to biofilm.

Plasma Components and Platelet Activation Are Essential for the Antimicrobial Properties of Autologous Platelet-Rich Plasma: An In Vitro Study

Autologous platelet concentrates are successfully adopted in a variety of medical fields to stimulate bone and soft tissue regeneration. The rationale for their use consists in the delivery of a wide range of platelet-derived bioactive molecules that promotes wound healing. In addition, antimicrobial properties of platelet concentrates have been pointed out. In this study, the effect of the platelet concentration, of the activation step and of the presence of plasmatic components on the antimicrobial activity of pure platelet-rich plasma was investigated against gram positive bacteria isolated from oral cavity. The antibacterial activity, evaluated as the minimum inhibitory concentration, was determined through the microdilution two-fold serial method. Results seem to suggest that the antimicrobial activity of platelet-rich plasma against Enterococcus faecalis, Streptococcus agalactiae, Streptococcus oralis and Staphylococcus aureus is sustained by a co-operation between plasma components and platelet-derived factors and that the activation of coagulation is a fundamental step. The findings of this study may have practical implications in the modality of application of platelet concentrates.

Treatment of atopic dermatitis eczema with a high concentration of Lactobacillus salivarius LS01 associated with an innovative gelling complex: a pilot study on adults.

Goals: To evaluate the efficacy of a highly concentrated Lactobacillus salivarius preparation containing a gelling complex formed by Streptococcus thermophilus ST10 and tara gum in the treatment of atopic dermatitis (AD). Background: Previous studies have demonstrated an improvement in AD symptoms after administration of the probiotic strain L. salivarius LS01. S. thermophilus ST10 and tara gum create a gelling complex that adheres to intestinal mucus and improves barrier function. Study: A prospective, controlled pilot trial was carried out to evaluate how the association of S. thermophilus ST10 and tara gum could improve the activity of L. salivarius LS01 administered at high doses to adults with AD. Twenty-five patients were included into the study: 13 were treated for 1 month with the active formulation, whereas 12 represented the placebo group. Scoring Atopic Dermatitis index was determined before and at the end of probiotic administration. Fecal samples were also collected to evaluate changes in bacterial counts of Staphylococcus aureus and clostridia. Results: A significant improvement in SCORAD index was observed in the probiotic group after 1 month of treatment, whereas no significant changes occurred in placebo patients. A slight decrease in fecal S. aureus count was observed in probiotic-treated patients. Conclusions: Data obtained in this study suggest a potential role for L. salivarius LS01 in the treatment of AD. The addition of tara gum and S. thermophilus ST10 seems to improve the overall efficacy of the probiotic strain, in particular shortening the time required for the onset of the positive effects. Further studies to investigate the activity of this preparation are advisable.

Role and antimicrobial resistance of staphylococci involved in prosthetic joint infections.

Purpose Staphylococci are responsible for approximately half of all prosthetic joint infections (PJIs) and they are often multi-drug resistant. The main purpose of this study was to evaluate the incidence of PJIs caused by staphylococci in our hospital from March 2010 to February 2012, with particular reference to antibiotic resistance in relation to their classification as contaminant or infecting isolates. Methods We analyzed samples recovered from 124 patients: most of them were male (55.8%) and the mean age was 66 ± 14 years. Prostheses derived from hip (54.8%) or knee (45.2%) replacement and they were processed by sonication. Isolates were identified using conventional biochemical methodologies. Antimicrobial susceptibility testing was carried out using the disk diffusion method as described by the European Committee on Antimicrobial Susceptibility Testing. Results A total of 135 staphylococci were isolated: the prevalent species was Staphylococcus aureus, but, on the whole, coagulase-negative staphylococci represented 57% of cases. Fifty-one isolates were recovered from a single sample and were therefore defined as contaminant. Linezolid and glycopeptides showed excellent activity versus all the tested isolates, while penicillin, levofloxacin, and erythromycin offered reduced activity against staphylococci. Interestingly, high resistance rates were observed for coagulase-negative staphylococci other than S. epidermidis classified as contaminant strains. Conclusions We observed a remarkable spread of coagulase-negative staphylococci as causative agents of PJIs; but most of them were classified as contaminants. However, because of their low susceptibility to the antibiotics tested, further studies are necessary to evaluate their role as pathogens or as true contaminants.

Biofilm Removal and Antimicrobial Activity of Two Different Air-Polishing Powders: An In Vitro Study

BACKGROUND Biofilm removal plays a central role in the prevention of periodontal and peri-implant diseases associated with microbial infections. Plaque debridement may be accomplished by air polishing using abrasive powders. In this study, a new formulation consisting of erythritol and chlorhexidine is compared with the standard glycine powder used in air-polishing devices. Their in vitro antimicrobial and antibiofilm effects on Staphylococcus aureus, Bacteroides fragilis, and Candida albicans are investigated. METHODS Biofilm was allowed to grow on sandblasted titanium disks and air polished with glycine or erythritol-chlorhexidine powders. A semiquantitative analysis of biofilm by spectrophotometric assay was performed. A qualitative analysis was also carried out by confocal laser scanning microscopy. Minimum inhibitory concentrations and minimum microbicidal concentrations were evaluated, together with the microbial recovery from the residual biofilm after air-polishing treatment. RESULTS The combination of erythritol and chlorhexidine displayed stronger antimicrobial and antibiofilm activity than glycine against all microbial strains tested. CONCLUSION Air polishing with erythritol-chlorhexidine seems to be a viable alternative to the traditional glycine treatment for biofilm removal.

Toll-Like Receptor 2 in Serum: a Potential Diagnostic Marker of Prosthetic Joint Infection?

ABSTRACT Prosthetic joint infection (PJI) is a severe complication of arthroplasty and is still lacking diagnostic gold standards. PJI patients display high Toll-like receptor 2 (TLR2) serum levels, correlating with canonical inflammatory markers (C-reactive protein [CRP], interleukin 6 [IL-6], tumor necrosis factor alpha [TNF-α], and IL-1). Therefore, TLR2 serum levels could be considered a new potential diagnostic tool in the early detection of PJI.