Treatment With Dithiothreitol Improves Bacterial Recovery From Tissue Samples in Osteoarticular and Joint Infections.

BACKGROUND: Prosthetic implants, periprosthetic and osteoarticular tissues are specimens of choice for diagnosis of bone and joint infections. Homogenization is considered the best procedure for treatment of tissues samples, but, it is not always performed in all laboratories. Dithiothreitol (DTT) has been proposed as an alternative treatment to sonication for microbiological diagnosis of prosthetic joint infections. In this study, the applicability of DTT treatment for processing of periprosthetic and osteoarticular tissues for diagnosis of bone and joint infections was evaluated and compared with normal saline solution treatment. METHODS: Periprosthetic tissue samples were collected from 70 consecutive patients (25 infected and 45 not infected). For each patient, samples from the same site were randomly allocated to DTT or saline treatment. Treated samples were centrifuged at 3000 rpm for 10 minutes. Aliquots from the concentrated samples were plated on agar plates and inoculated in broths. Sensitivity and specificity were calculated for each treatment. RESULTS: Microbial growth was observed in samples from 14 and 11 infected patients after DTT and saline treatments, respectively. Concordance between the 2 methods was observed in the 85.7% of cases. Sensitivity was 88.0% for DTT and 72.0% for saline. Specificity was 97.8% and 91.1% for DTT and saline, respectively. Treatment with DTT showed higher sensitivity and specificity with respect to the method routinely used in our laboratory. CONCLUSION: DTT treatment may be considered a practicable strategy for microbiological analysis of tissues for diagnosis of bone and joint infections.

May osteoarticular infections be influenced by vitamin D status? An observational study on selected patients.

BACKGROUND: Vitamin D deficiency has been associated with a high number of health outcomes, and its role on the immune system has been deeply investigated in recent years, although poor data are still available on vitamin D status in orthopedic infections including those of prosthetic implants. METHODS: We focused on preoperative values of 25(OH)D in selected groups of patients with septic (Group A) or aseptic (Group B) prosthetic loosening, infective bone disease such as septic arthritis and osteomyelitis (Group C) and other orthopedic pathologies (Group D) to evaluate differences in the vitamin D status. RESULTS: A high prevalence of vitamin D deficiency was recorded among the study population (16.5 ± 5.4 ng/mL, mean ± SD). Interestingly, all patients with an infection presented a higher 25(OH)D concentration (17.7 ± 5.3 ng/mL) in respect to uninfected ones (15.1 ± 5.6 ng/mL). Significantly higher levels of 25(OH)D were observed in patients with prosthetic joint infection (18.5 ± 6.5 ng/mL), when compared with those presenting an aseptic loosening (13.6 ± 9.4 ng/mL). CONCLUSIONS: Deficiency in vitamin D levels have been found in orthopaedic patients. Prosthetic joint infections seems to be associated to higher values of vitamin D in respect to other bone infections or to other orthopaedic conditions requiring surgery. More studies are needed to improve the knowledge on vitamin D status in these patients and to better clarify the role of vitamin D in relation to osteoarticular infections.

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.

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.

Dynamic QuantiFERON Response in Psoriasis Patients Taking Long-Term Biologic Therapy.

INTRODUCTION: The risk of active tuberculosis is increased in psoriasis patients receiving biologic drug therapy. The QuantiFERON-TB Gold In-Tube assay (QFT) is used for latent tuberculosis screening in these patients. This study presents a retrospective analysis on repeated QFT assays, investigating the influence of biologic drugs and isoniazid therapy on the outcome of the assay. METHODS: Serial QFTs of 58 psoriasis patients, who received biologic drug therapy, were evaluated at baseline and after 12 months of treatment. Patients were retrospectively divided in four groups according to QFT results at baseline and at follow-up: patients having a QFT reversion (from positive to negative results); patients with a conversion (from negative to positive); patients confirming the baseline results, either positive or negative. RESULTS: At the end of the 12-months period, 11.1% of patients with a negative QFT result at baseline presented a conversion, showing low interferon (IFN)-gamma values, whereas 6.9% of positive patients presented a QFT reversion. When the test was repeated after 2-3 months without isoniazid chemoprophylaxis, patients with QFT conversion showed negative results. No patient developed active tuberculosis. CONCLUSIONS: In patients undergoing biologic therapy, a positive QFT assay needs to be further confirmed, as false-positive results may occur after long-term therapy. Repeating QFT tests in patients with low IFN-gamma values could reduce the incidence of false-positive latent tuberculosis infection diagnosis, thus preventing unnecessary tuberculosis chemoprophylaxis. In conclusion, a dynamic QFT response is possible in psoriasis patients undergoing biologic therapy.

Does dithiothreitol improve bacterial detection from infected prostheses? A pilot study.

BACKGROUND: Sonication and scraping of infected prostheses usually are used to improve diagnosis of prosthetic infections, reducing false negatives. Chemical methods that reduce biofilms also may allow higher levels of detection. QUESTIONS/PURPOSES: We therefore asked: (1) Do dithiothreitol (DTT) and N-acetylcysteine (NAC) remove bacteria from biofilm formed on prosthetic materials? (2) Is bacterial recovery affected by differing DTT and NAC concentrations and incubation times? (3) Do treatments with DTT and NAC detach the same amounts of bacteria from biofilm on prosthetic materials as sonication and scraping? (4) Are these methods reproducible? METHODS: We treated polyethylene and titanium discs covered by biofilm formed by Pseudomonas aeruginosa and Staphylococcus aureus with DTT or NAC solutions at different concentrations for different times. We compared colony counts of S aureus, P aeruginosa, Staphylococcus epidermidis and Escherichia coli after treatment with NAC, DTT, sonication and scraping. We determined colony counts after treatment of biofilm formed by one strain of S aureus and one of P aeruginosa on five discs of each material analyzed on the same day and on five discs analyzed on five consecutive days. RESULTS: Mean colony counts (LogCFU/mL) obtained after treatment with 1 g/L DTT for 15 minutes (5.3) were similar to those after sonication (4.9) and greater than those obtaining by scraping (3.4) and treatment with 2 g/L NAC for 30 minutes (1.9). DTT and sonication showed good reproducibility. CLINICAL RELEVANCE: Our data suggest that treatment of prostheses with DTT may be a reasonable alternative to sonication to improve detection of biofilm-associated bacteria and supplement conventional laboratory culturing techniques for diagnosing periprosthetic infections.