Primary deep infection following joint replacement surgery accounts for 7% of all revisions. Glass polyalkenoate cements (GPCs) have previously been shown to exhibit antibacterial properties. The present study had two objectives. The first was to determine if addition of tri-sodium citrate (TSC) to the powder phase of an Al-free GPC (0.04 SrO-0.12 CaO-0.36 ZnO-0.48 SiO2, by mole fraction) enhanced the resultant cement's antibacterial properties against three strains of bacteria that are commonly found in periprosthetic sites following total joint replacements (TJRs); namely, E. coli, B. fragilis, and S. epidermidis. Four cement sets were prepared, which contained 0 wt% TSC (control), 5 wt% TSC, 10 wt% TSC, and 15 wt% TSC. All the TSC-modified cements were found to exhibit large inhibition zones against all the bacterial strains, especially the cement containing 15 wt% TSC against E. coli. The antibacterial properties of the TSC containing GPCs are attributed to the release of Zn and Na ions from the cements and the presence of the TSC. The second objective was to investigate if, when a modified GPC is embedded in a bovine bone model, ionic transfer occurs. It was found that Zn ions migrated from the cement to the surrounding bone, particularly at the cement-bone interface. This is a desirable outcome as Zn ions are known to play a vital role in both bone metabolism and the regeneration of healthy bone. The present results point to the potential clinical benefits of using TSC-modified GPCs in TJRs.
BackgroundThe C10 family of cysteine proteases includes enzymes that contribute to the virulence of bacterial pathogens, such as SpeB in Streptococcus pyogenes. The presence of homologues of cysteine protease genes in human commensal organisms has not been examined. Bacteroides fragilis is a member of the dominant Bacteroidetes phylum of the human intestinal microbiota, and is a significant opportunistic pathogen.ResultsFour homologues of the streptococcal virulence factor SpeB were identified in the B. fragilis genome. These four protease genes, two were directly contiguous to open reading frames predicted to encode staphostatin-like inhibitors, with which the protease genes were co-transcribed. Two of these protease genes are unique to B. fragilis 638R and are associated with two large genomic insertions. Gene annotation indicated that one of these insertions was a conjugative Tn-like element and the other was a prophage-like element, which was shown to be capable of excision. Homologues of the B. fragilis C10 protease genes were present in a panel of clinical isolates, and in DNA extracted from normal human faecal microbiota.ConclusionsThis study suggests a mechanism for the evolution and dissemination of an important class of protease in major members of the normal human microbiota.
BackgroundBacteroides fragilis and Bacteroides thetaiotaomicron are members of the normal human intestinal microbiota. However, both organisms are capable of causing opportunistic infections, during which the environmental conditions to which the bacteria are exposed change dramatically. To further explore their potential for contributing to infection, we have characterized the expression in B. thetaiotaomicron of four homologues of the gene encoding the C10 cysteine protease SpeB, a potent extracellular virulence factor produced by Streptococcus pyogenes.ResultsWe identified a paralogous set of genes (btp genes) in the B. thetaiotaomicron genome, that were related to C10 protease genes we recently identified in B. fragilis. Similar to C10 proteases found in B. fragilis, three of the B. thetaiotaomicron homologues were transcriptionally coupled to genes encoding small proteins that are similar in structural architecture to Staphostatins, protease inhibitors associated with Staphopains in Staphylococcus aureus. The expression of genes for these C10 proteases in both B. fragilis and B. thetaiotaomicron was found to be regulated by environmental stimuli, in particular by exposure to oxygen, which may be important for their contribution to the development of opportunistic infections.ConclusionsGenes encoding C10 proteases are increasingly identified in operons which also contain genes encoding proteins homologous to protease inhibitors. The Bacteroides C10 protease gene expression levels are responsive to different environmental stimuli suggesting they may have distinct roles in the bacterial-host interaction.
Infection following surgery can result in significant pain and morbidity for patients undergoing vertebroplasty/kyphoplasty, and often results in revision surgery. This study focuses on the development of Al-free glass polyalkenoate cements (GPCs) based on 0.04SrO-0.12CaO-0.36ZnO-0.48SiO( 2) glass, with the intent of optimizing their antibacterial efficacy by incorporating low-molecular-weight polyacrylic acids (PAA) and trisodium citrate (TSC), and evaluating the resultant GPCs against bacteria relevant to spinal infections, P. aeruginosa and E. coli. Ion-release profiles were determined for the GPC formulation containing E6 PAA (Cement A) and E7 PAA (Cement B), and Zn, Na, and Sr release was recorded over 1, 7, and 30 days. Inhibition was found in E. coli at each time period (0-30 days) and this generally decreased with exposure time in water. The largest GPC inhibition zones were produced by Cement A (6 mm); however the control material Simplex P + tobramycin produced much higher inhibition zones (11 mm). When testing the GPC against P. aeruginosa, inhibition was only present at the 0-day time period. Simplex P + tobramycin was found to produce inhibition at each time frame. Analysis of the agar from the inhibition zone of the E. coli test revealed that there is a significant change in Zn concentration as compared to a control agar specimen, which suggests that Zn release is responsible for the antibacterial effect of the GPCs.
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