Early outcome of culture-negative infection in open fractures of the lower limb: A prospective study.

Background: Culture-negative infections in open long bone fractures are frequently encountered in clinical practice. We aimed to identify the rate and outcome of culture-negative infections in open long bone fractures of lower limb. Methodology: A prospective cohort study was conducted from November 2015 to May 2017 on Gustilo and Anderson Grade III open long bone fractures of the lower limb. Demographic data, injury details, time from injury to receiving antibiotics and index surgical procedure were noted. Length of hospital stay, number of additional surgeries and occurrence of complications were also noted. Patients with infected open fractures were grouped as culture positive or culture negative depending on the isolation of infecting microorganisms in deep intraoperative specimen. The clinical outcome of these two groups was statistically analysed. Results: A total of 231 patients with 275 open fractures involving the femur, tibia or fibula were studied. There was clinical signs of infection in 84 patients (36.4%) with 99 fractures (36%). Forty-three patients (51.2%) had positive cultures and remaining 41 patients had negative cultures (48.8%). The rate of culture-negative infection in open type III long bone fractures in our study was 17.7%. There was no statistical difference in the clinical outcome between culture-negative and culture-positive infections. Conclusion: Failure to identify an infective microorganism in the presence of clinical signs of infection is routinely seen in open fractures and needs to be treated aggressively.

Comparative Genomics Study of Staphylococcus epidermidis Isolates from Orthopedic-Device-Related Infections Correlated with Patient Outcome.

Staphylococcus epidermidis has emerged as an important opportunistic pathogen causing orthopedic-device-related infections (ODRI). This study investigated the association of genome variation and phenotypic features of the infecting S. epidermidis isolate with the clinical outcome for the infected patient. S. epidermidis isolates were collected from 104 patients with ODRI. Their clinical outcomes were evaluated, after an average of 26 months, as either "cured" or "not cured." The isolates were tested for antibiotic susceptibility and biofilm formation. Whole-genome sequencing was performed on all isolates, and genomic variation was related to features associated with "cured" and "not cured." Strong biofilm formation and aminoglycoside resistance were associated with a "not-cured" outcome (P = 0.031 and P < 0.001, respectively). Based on gene-by-gene analysis, some accessory genes were more prevalent in isolates from the "not-cured" group. These included the biofilm-associated bhp gene, the antiseptic resistance qacA gene, the cassette chromosome recombinase-encoding genes ccrA and ccrB, and the IS256-like transposase gene. This study identifies biofilm formation and antibiotic resistance as associated with poor outcome in S. epidermidis ODRI. Whole-genome sequencing identified specific genes associated with a "not-cured" outcome that should be validated in future studies. (The study has been registered at ClinicalTrials.gov with identifier NCT02640937.).

Low-level laser therapy to the mouse femur enhances the fungicidal response of neutrophils against Paracoccidioides brasiliensis.

Neutrophils (PMN) play a central role in host defense against the neglected fungal infection paracoccidioidomycosis (PCM), which is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). PCM is of major importance, especially in Latin America, and its treatment relies on the use of antifungal drugs. However, the course of treatment is lengthy, leading to side effects and even development of fungal resistance. The goal of the study was to use low-level laser therapy (LLLT) to stimulate PMN to fight Pb in vivo. Swiss mice with subcutaneous air pouches were inoculated with a virulent strain of Pb or fungal cell wall components (Zymosan), and then received LLLT (780 nm; 50 mW; 12.5 J/cm2; 30 seconds per point, giving a total energy of 0.5 J per point) on alternate days at two points on each hind leg. The aim was to reach the bone marrow in the femur with light. Non-irradiated animals were used as controls. The number and viability of the PMN that migrated to the inoculation site was assessed, as well as their ability to synthesize proteins, produce reactive oxygen species (ROS) and their fungicidal activity. The highly pure PMN populations obtained after 10 days of infection were also subsequently cultured in the presence of Pb for trials of protein production, evaluation of mitochondrial activity, ROS production and quantification of viable fungi growth. PMN from mice that received LLLT were more active metabolically, had higher fungicidal activity against Pb in vivo and also in vitro. The kinetics of neutrophil protein production also correlated with a more activated state. LLLT may be a safe and non-invasive approach to deal with PCM infection.

Efficacy of ciprofloxacin implants in treating experimental osteomyelitis.

Ciprofloxacin (CFX) implants containing poly(D,L-lactide) and calcium phosphates (tricalcium phosphate and hydroxyapatite) was evaluated in 50 rabbits in an experimental osteomyelitis model. Their femoral cavity was inoculated with Staphylococcus aureus. After 2 weeks, the infected focus was cleaned out and the delivery system implanted. The infection and subsequent response to treatment were evaluated by microbiological analysis, biochemical and hematological markers, body weight, temperature, clinical signs, X-rays, and histology. Infected bone cultures, treated with CFX implants, showed reduced bacterial growth against controls. All CFX was released within 6 weeks. All animals recovered within 4 weeks. Even 12 weeks after implantation, no recurrence of infection was observed. Serum C-reactive protein, platelet, and leukocyte levels increased in all animals before treatment, and 4 weeks after it were maintained or rose in control animals, while decreased to normal levels in treated ones. Body weight was characterized by pretreatment losses, then gains during recuperation, or further loss in untreated animals; with no significant intraindividual differences in body temperature. Body weight, leucocytes, platelets, and C-reactive protein turned out to be highly useful markers for monitoring this kind of infection and its treatment. CFX implants demonstrated to be an effective therapy for S. aureus bone infection. Their efficacy was also reflected in decreasing severity of clinical signs, nonprogress of radiological signs indicative of infection, and good integration into bone structure. Histological examination revealed repair, with new bone formation extending into implants.

Moxifloxacin superior to vancomycin for treatment of bone infections--a study in rats.

BACKGROUND: Increasing resistance rates towards conventional antibiotics necessitate investigations of the efficacy of newly developed antibiotics. Thus, in a rat study, we compared the efficacy of moxifloxacin and vancomycin in the treatment of a local Staphylococcus aureus bone infection. METHOD: The femoral medullary cavities of 36 Wistar rats were contaminated with 100 muL of an oxacillin-sensitive Staphylococcus aureus strain (ATCC 29213) at 10(8) cfu/mL. On the seventh day, antibiotic treatment with moxifloxacin (10 mg/kg twice daily i.p.) or vancomycin (15 mg/kg twice daily i.p.) was commenced in 12 animals each. 12 control animals were left untreated. After 21 days, the infected femurs were explanted and the bacterial counts (cfu/g) were determined. RESULTS: In the control group, a median of 3.42 x 10(6) cfu/g (LQ/UQ 1.09 x 10(6)/ 1.55 x 10(7)) was cultured, with a median of 2.53 x 10(6) cfu/g (LQ/UQ 1.95 x 10(6)/ 4.25 x 10(6)) in the vancomycin group and a median of 2.49 x 10(5) cfu/g (LQ/UQ 2.84 x 10(4)/ 3.75 x 10(5)) in the moxifloxacin group. The bacterial count was reduced by treatment with moxifloxacin both in comparison with the control group (p < 0.001), and in comparison with treatment with vancomycin (p < 0.001). There was no statistically significant difference between the vancomycin group and the control group (p = 0.53). INTERPRETATION: In contrast to vancomycin, moxifloxacin proved to be an effective antibiotic for the treatment of bone infections due to Staphylococcus aureus in our animal model.

In vivo evaluation of teicoplanin- and calcium sulfate-loaded PMMA bone cement in preventing implant-related osteomyelitis in rats.

The objective of this study was to evaluate the efficacy of teicoplanin- and calcium sulphate-loaded polymethylmethacrylate (PMMA) bone cements in preventing experimental implant-related osteomyelitis in rats. Four groups of antibiotic-loaded rods were prepared and were implanted into the lateral condylus of the rat femur after inoculation of Staphylococcus aureus. The effectiveness of these were assessed microbiologically, radiographically, and histopathologically. Radiographic evaluation revealed a significant reduction of periostal reaction and osteolysis in rats that received calcium sulphate- and teicoplanin-loaded rods. Histopathological evaluation confirmed these results. Acute infection and bone necrosis were found to be significantly lower in rats that had received calcium sulphate- and teicoplanin-loaded rods. The addition of calcium sulfate to teicoplanin-loaded PMMA bone cement appeared satisfactory as an antibiotic-carrying system for prophylaxis of experimental implant-related osteomyelitis, but further investigations are needed to reach definitive statements for clinical applications.

Prevention of infection with tobramycin-containing bone cement or systemic cefazolin in an animal model.

We investigated in an animal model the efficacy of tobramycin-containing bone cement and systemic cefazolin for infection prophylaxis. In 18 female rabbits, the femoral cavity was inoculated with Staphylococcus aureus before injection of bone cement. The first group of six rabbits received tobramycin-containing Simplex-P bone cement. Two other groups of six rabbits received plain Simplex-P bone cement. Preoperatively, in one of the two latter groups cefazolin was administered intravenously. The other group served as untreated controls. The rabbits were monitored for clinical signs of infection. At 7 days' follow-up, the femora were harvested and cultures from the bone adjacent to the cement plug were quantified. Cultures from the rabbits which received antibiotic prophylaxis (either cefazolin systemically or tobramycin-containing bone cement) were all negative. In contrast, all rabbits in the untreated control group had positive cultures. These rabbits also had other signs of infection such as an elevated erythrocyte sedimentation rate and loss of body weight. Culture results were confirmed by the absence of bacterial DNA in the polymerase chain reaction hybridization assay. In conclusion, we found that both tobramycin-containing bone cement and systemic cefazolin are effective in preventing implant bed infection in rabbits up to 7 days after contamination with S. aureus.

Prophylaxis of implant-related staphylococcal infections using tobramycin-containing bone cement.

In a rabbit model, premixed tobramycin-containing bone cement was studied for its efficacy to prevent infections with two frequently encountered staphylococcal species in arthroplasty surgery. After intramedullary inoculation with staphylococci, either standard or premixed tobramycin-containing Simplex-P bone cement was injected in the right femur of 120 rabbits. Development of infection was examined by culture of femoral bone after 7 or 28 days. Loss of body weight and elevated erythrocyte sedimentation rate in the control rabbits inoculated with Staphylococcus aureus were seen in the first postoperative week, returning to normal in 28 days. Inoculation with Staphylococcus epidermidis resulted only in a low-grade infection. All rabbits receiving premixed tobramycin-containing bone cement were free of signs of infection, and all their cultures were negative. Culture yield from Staphylococcus aureus controls increased with time and inoculum dose. Staphylococcus epidermidis controls needed higher inoculum doses to establish an infection, while culture yield decreased in time. These differences in mode of prosthesis-related infection are explained by differences in virulence factors.

In vitro and in vivo release of ciprofloxacin from PLGA 50:50 implants.

Poly(lactides-co-glycolides) [PLGA] are widely investigated biodegradable polymers and are extensively used in several biomaterials applications as well as drug delivery systems. These polymers degrade by bulk hydrolysis of ester bonds and break down into their constituent monomers, lactic and glycolic acids which are excreted from the body. The purpose of this investigation was to develop and characterize a biodegradable, implantable delivery system containing ciprofloxacin hydrochloride (HCl) for the localized treatment of osteomyelitis and to study the extent of drug penetration from the site of implantation into the bone. Osteomyelitis is an inflammatory bone disease caused by pyogenic bacteria and involves the medullary cavity, cortex and periosteum. The advantages of localized biodegradable therapy include high, local antibiotic concentration at the site of infection, as well as, obviation of the need for removal of the implant after treatment. PLGA 50:50 implants were compressed from microcapsules prepared by nonsolvent-induced phase-separation using two solvent-nonsolvent systems, viz., methylene chloride-hexane (non-polar) and acetone-phosphate buffer (polar). In vitro dissolution studies were performed to study the effect of manufacturing procedure, drug loading and pH on the release of ciprofloxacin HCl. The extent of penetration of the drug from the site of implantation was studied using a rabbit model. The results of in vitro studies illustrated that drug release from implants made by the nonpolar method was more rapid as compared to implants made by the polar method. The release of ciprofloxacin HCl. The extent of the penetration of the drug from the site of implantation was studied using a rabbit model. The results of in vitro studies illustrated that drug release from implants made by the nonpolar method was more rapid as compared to implants made by the polar method. The release of ciprofloxacin HCl from the implants was biphasic at < or = 20% w/w drug loading, and monophasic at drug loading levels > or = 35% w/w. In vivo studies indicated that PLGA 50:50 implants were almost completely resorbed within five to six weeks. Sustained drug levels, greater than the minimum inhibitory concentration (MIC) of ciprofloxacin, up to 70 mm from the site of implantation, were detected for a period of six weeks.

The use of bacitracin irrigation to prevent infection in postoperative skeletal wounds. An experimental study.

In dogs, irrigation of contaminated osseous wounds with bacitracin eliminated clinical evidence of infection and significantly reduced the number of positive cultures and pathological evidence of infection when compared with dogs that received no treatment or irrigation with normal saline solution. The inocula contained more organisms than are introduced into a wound during an elective orthopaedic operation. The use of bacitracin in the prevention of postoperative Staphylococcus aureus infection of bone in humans may be justified.