Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 305
Filtrar
1.
Microbiol Spectr ; 9(2): e0026721, 2021 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-34523997

RESUMO

Rapid synovial fluid-induced aggregation of Staphylococcus aureus is currently being investigated as an important factor in the establishment of periprosthetic joint infections (PJIs). Pathogenic advantages of aggregate formation have been well documented in vitro, including recalcitrance to antibiotics and protection from host immune defenses. The objective of the present work was to determine the strain dependency of synovial fluid-induced aggregation by measuring the degree of aggregation of 21 clinical S. aureus isolates cultured from either PJI or bloodstream infections using imaging and flow cytometry. Furthermore, by measuring attached bacterial biomass using a conventional crystal violet assay, we assessed whether there is a correlation between the aggregative phenotype and surface-associated biofilm formation. While all of the isolates were stimulated to aggregate upon exposure to bovine synovial fluid (BSF) and human serum (HS), the extent of aggregation was highly variable between individual strains. Interestingly, the PJI isolates aggregated significantly more upon BSF exposure than those isolated from bloodstream infections. While we were able to stimulate biofilm formation with all of the isolates in growth medium, supplementation with either synovial fluid or human serum inhibited bacterial surface attachment over a 24 h incubation. Surprisingly, there was no correlation between the degree of synovial fluid-induced aggregation and quantity of surface-associated biofilm as measured by a conventional biofilm assay without host fluid supplementation. Taken together, our findings suggest that synovial fluid-induced aggregation appears to be widespread among S. aureus strains and mechanistically independent of biofilm formation. IMPORTANCE Bacterial infections of hip and knee implants are rare but devastating complications of orthopedic surgery. Despite a widespread appreciation of the considerable financial, physical, and emotional burden associated with the development of a prosthetic joint infection, the establishment of bacteria in the synovial joint remains poorly understood. It has been shown that immediately upon exposure to synovial fluid, the viscous fluid in the joint, Staphylococcus aureus rapidly forms aggregates which are resistant to antibiotics and host immune cell clearance. The bacterial virulence associated with aggregate formation is likely a step in the establishment of prosthetic joint infection, and as such, it has the potential to be a potent target of prevention. We hope that this work contributes to the future development of therapeutics targeting synovial fluid-induced aggregation to better prevent and treat these infections.


Assuntos
Aderência Bacteriana/fisiologia , Biofilmes/crescimento & desenvolvimento , Infecções Relacionadas à Prótese/microbiologia , Staphylococcus aureus/crescimento & desenvolvimento , Líquido Sinovial/microbiologia , Animais , Bovinos , Prótese de Quadril/microbiologia , Humanos , Prótese do Joelho/microbiologia , Soro/microbiologia , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação , Membrana Sinovial/microbiologia
2.
Nagoya J Med Sci ; 83(1): 151-157, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33727746

RESUMO

In recent years, Cutibacterium acnes (C. acnes) has been reported to affect postoperative outcomes. The purpose of this study was to examine the detection rate and clinical features of C. acnes infection after open shoulder surgery. Fifty-nine patients (33 males and 26 females; mean age, 69.1 years) were included. Samples were collected from a skin swab at the incision site prior to skin preparation. Further samples were collected from synovial swabs at the glenohumeral joint immediately after incision and before incision closure. Samples with C. acnes-positive skin swab cultures were defined as Group A, and those with negative cultures were defined as Group N. Age, sex, presence of diabetes mellitus, operation time, presence of deep infection after surgery, and rate of positive synovial swab cultures were compared between groups. There were 27 patients in Group A (mean age 69.1±13.3 [SD], 21 males and 6 females) and 32 patients in Group N (mean age 69.1±11.0 [SD], 12 males and 20 females). No significant difference in the presence of diabetes mellitus and operation time were found between groups. From the glenohumeral joint immediately after incision, C. acnes was detected in 22.2% and 0% of patients in Group A and Group N, respectively. For the glenohumeral joint before incision closure, C. acnes was detected in 22.2% and 0% of patients in Group A and Group N, respectively, demonstrating a significantly higher rate in Group A. Our findings suggest that the route of infection following open shoulder surgery is via contamination.


Assuntos
Infecções por Bactérias Gram-Positivas/epidemiologia , Propionibacterium acnes/isolamento & purificação , Articulação do Ombro/microbiologia , Pele/microbiologia , Membrana Sinovial/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Artroplastia do Ombro , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/epidemiologia , Lesões do Manguito Rotador/cirurgia , Ombro/cirurgia
3.
PLoS One ; 15(12): e0242868, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33382721

RESUMO

Rheumatoid arthritis (RA) and periodontitis (PD) are chronic inflammatory diseases that appear to occur in tandem. However, the mutual impact PD exerts on RA and vice versa has not yet been defined. To address this issue, we set up an animal model and analyzed how two prime inducers of periodontitis-Porphyromonas gingivalis (Pg) and Aggregatibacter actinomycetemcomitans (Aa)-differ in their pathogenic potential. Our experimental setup included collagen induced arthritis (CIA) in the mouse, oral inoculation with Pg or Aa to induce alveolar bone loss and the combination of both diseases in inverted orders of events. Neither pathobiont impacted on macroscopic arthritis and arthritis did not exacerbate alveolar bone loss. However, there were subtle differences between Pg and Aa with the former inducing more alveolar bone loss if PD was induced before CIA. On a molecular level, Pg and Aa led to differential expression patterns in the synovial membranes that were reminiscent of cellular and humoral immune responses, respectively. The Pg and Aa specific signatures in the synovial proteomes suggest a role for oral pathogens in shaping disease subtypes and setting the stage for subsequent therapy response.


Assuntos
Aggregatibacter actinomycetemcomitans/fisiologia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/microbiologia , Porphyromonas gingivalis/fisiologia , Proteoma/metabolismo , Membrana Sinovial/metabolismo , Animais , Citocinas/metabolismo , Camundongos , Periodontite/microbiologia , Membrana Sinovial/microbiologia
4.
Int J Mol Sci ; 21(22)2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-33207573

RESUMO

Osteoarthritis (OA) is the most common joint disorder in the United States, and the gut microbiome has recently emerged as a potential etiologic factor in OA development. Recent studies have shown that a microbiome is present at joint synovia. Therefore, we aimed to characterize the intra-articular microbiome within osteoarthritic synovia and to illustrate its role in OA disease progression. RNA-sequencing data from OA patient synovial tissue was aligned to a library of microbial reference genomes to identify microbial reads indicative of microbial abundance. Microbial abundance data of OA and normal samples was compared to identify differentially abundant microbes. We computationally explored the correlation of differentially abundant microbes to immunological gene signatures, immune signaling pathways, and immune cell infiltration. We found that microbes correlated to OA are related to dysregulation of two main functional pathways: increased inflammation-induced extracellular matrix remodeling and decreased cell signaling pathways crucial for joint and immune function. We also confirmed that the differentially abundant and biologically relevant microbes we had identified were not contaminants. Collectively, our findings contribute to the understanding of the human microbiome, well-known OA risk factors, and the role microbes play in OA pathogenesis. In conclusion, we present previously undiscovered microbes implicated in the OA disease progression that may be useful for future treatment purposes.


Assuntos
Bactérias , Articulação do Joelho/microbiologia , Microbiota , Osteoartrite do Joelho/microbiologia , Membrana Sinovial/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Humanos , RNA-Seq
5.
BMC Infect Dis ; 20(1): 672, 2020 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-32938418

RESUMO

BACKGROUND: Mycobacterium jacuzzii (M. jacuzzii) was first isolated in 2003 by insertion of breast implants in Tel Aviv, Israel. In this case report, we describe our experience in detection of M. jacuzzii using phenotypic and genotypic test of wrist synovial sample. CASE PRESENTATION: A 73-year-old woman complained of pain and swelling in the right wrist for 4 months. Her body temperature was 37-38 °C, and symptoms, such as pain, swelling, and some movement limitation, were reported. Clinical laboratory parameters showed an elevated C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR), and white blood cells (WBC) count. The sequences of hsp65, rpoB, 16S rDNA, and sodA genes indicated very high homology to M. jacuzzii. CONCLUSION: We report a case of synovial infection caused by M. jacuzzii in a patient with severe wrist pain in Iran, who was treated with amikacin, levofloxacin, and ethambutol. The outcomes of treatment after 8 months were positive, and no recurrence of infection was reported in the patient.


Assuntos
Implantes de Mama/efeitos adversos , Infecções por Mycobacterium/diagnóstico , Mycobacterium/genética , Membrana Sinovial/microbiologia , Idoso , Amicacina/uso terapêutico , Antibacterianos/uso terapêutico , Sedimentação Sanguínea , Feminino , Humanos , Irã (Geográfico) , Contagem de Leucócitos , Mycobacterium/classificação , Mycobacterium/isolamento & purificação , Infecções por Mycobacterium/tratamento farmacológico , Infecções por Mycobacterium/microbiologia , Filogenia , RNA Ribossômico 16S/classificação , RNA Ribossômico 16S/metabolismo , Punho/microbiologia
6.
Vet Immunol Immunopathol ; 227: 110057, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32554268

RESUMO

Mycoplasma bovis causes chronic arthritis in calves, presenting as osteolysis in affected joints. Matrix metalloproteinase-3 (MMP-3), an enzyme involved in cartilage degradation, is produced by synovial cells. Production of this proteinase is regulated by interleukin (IL)-1ß, which is produced by mononuclear cells. Both factors are known to play important roles in osteolysis in human autoimmune and bacterial arthritis. However, the pathophysiology of Mycoplasma arthritis (MA) has not been elucidated. In this study, we evaluated the levels of MMP-3 and IL-1ß in synovial fluid (SF) from MA calves and examined the effect of IL-1ß on MMP-3 expression in bovine synovial cells in vitro. Levels of MMP-3 and IL-1ß in SF from MA calves were significantly higher than those of clinically healthy calves. Mycoplasma bovis induced significant increases in the expression of IL-1ß mRNA and protein in mononuclear cells, compared with cells not exposed to M. bovis. Interestingly, the supernatant of mononuclear cells stimulated with M. bovis contained high levels of IL-1ß, which induced higher expression of MMP-3 mRNA and protein in synovial cells than direct stimulation by M. bovis. Recombinant bovine IL-1ß also induced increased MMP-3 mRNA and protein expression in synovial cells. Our results indicate that M. bovis induces IL-1ß expression by bovine mononuclear cells, and this cytokine then promotes MMP-3 production by synovial cells. These findings suggest that MMP-3 and IL-1ß are key factors in the development of osteolysis in MA calves.


Assuntos
Artrite Infecciosa/veterinária , Doenças dos Bovinos/imunologia , Interleucina-1beta/imunologia , Metaloproteinase 3 da Matriz/genética , Infecções por Mycoplasma/veterinária , Membrana Sinovial/imunologia , Animais , Artrite Infecciosa/imunologia , Bovinos , Doenças dos Bovinos/microbiologia , Células Cultivadas , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/microbiologia , Infecções por Mycoplasma/imunologia , Mycoplasma bovis , Líquido Sinovial/química , Líquido Sinovial/imunologia , Membrana Sinovial/citologia , Membrana Sinovial/microbiologia , Regulação para Cima
8.
J Shoulder Elbow Surg ; 29(1): 20-26, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31619355

RESUMO

BACKGROUND: The utility of next-generation sequencing (NGS) in differentiating between active infection and contaminant or baseline flora remains unclear. The purpose of this study is to compare NGS with culture-based methods in primary shoulder arthroplasty. METHODS: A prospective series of primary shoulder arthroplasty patients with no history of infection or antibiotic use within 60 days of surgery was enrolled. All patients received standard perioperative antibiotics. After skin incision, a 10 × 3-mm sample of the medial skin edge was excised. A 2 × 2-cm synovial tissue biopsy was taken from the rotator interval after subscapularis takedown. Each sample set was halved and sent for NGS and standard cultures. RESULTS: Samples from 25 patients were analyzed. Standard aerobic/anaerobic cultures were positive in 10 skin samples (40%, 95% confidence interval [CI] 20%-60%) and 3 deep tissue samples (12%, 90% CI 1%-23%]). NGS detected ≥1 bacterial species in 17 of the skin samples (68%, 95% CI 49%-87%) and 7 deep tissue samples (28%, 95% CI 9%-47%). There was a significant difference (P < .03) in the mean number of bacterial species detected with NGS between the positive standard culture (1.6 species) and the negative standard culture groups (5.7 species). CONCLUSION: NGS identified bacteria at higher rates in skin and deep tissue samples than standard culture did in native, uninfected patients undergoing primary procedures. Further research is needed to determine which NGS results are clinically relevant and which are false positives before NGS can be reliably used in orthopedic cases.


Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/diagnóstico , Sequenciamento de Nucleotídeos em Larga Escala , Articulação do Ombro/cirurgia , Pele/microbiologia , Membrana Sinovial/microbiologia , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Artroplastia , Infecções Bacterianas/microbiologia , Contagem de Colônia Microbiana , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos
9.
Arthroscopy ; 35(9): 2571-2577, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31351811

RESUMO

PURPOSE: To analyze the utility of arthroscopic biopsies for detection of periprosthetic infection in painful shoulder arthroplasty without objective signs of infection. METHODS: A retrospective analysis of all patients who underwent a diagnostic arthroscopy for painful shoulder arthroplasty from June 2012 through July 2018 was performed. Patients with a subsequent revision shoulder arthroplasty after diagnostic arthroscopy were included. Arthroscopic tissue culture results were compared with the culture results of intraoperative tissue samples obtained at the time of open revision surgery. A minimum of 3 tissue samples from synovia and bone-prosthesis interface with signs of synovitis or abnormal appearance was routinely collected. Cases with 2 or more positive cultures for the same microorganism obtained at open revision surgery were considered as true presence of infection. The study protocol was reviewed and approved by the institutional ethics committee. RESULTS: Twenty-three cases in 22 patients were included in this study. Five of these 23 cases were classified as true infection based on the samples obtained during open revision surgery, and 16 cases had a positive culture in diagnostic arthroscopy. Cutibacterium acnes was isolated in each case. Classifying any microbiologic growth in the arthroscopic biopsies as positive resulted in a sensitivity and negative predictive value of 100%, specificity of 39%, and positive predictive value of 31.3% for the detection of a periprosthetic shoulder infection (PPSI). If at least 2 positive samples with the same microbiologic growth in the arthroscopic biopsies were considered as positive, sensitivity and negative predictive value dropped to 80% and 94.4%, respectively, but the specificity and positive predictive value increased to 94.4% and 80%, respectively. CONCLUSIONS: Diagnostic arthroscopy is a useful diagnostic tool in patients with suspicion but no clear evidence of PPSI. Arthroscopically obtained tissue biopsies for culture offer a high sensitivity and specificity in the diagnosis of PPSI if at least 2 cultures positive for the same microorganism are considered as infection. LEVEL OF EVIDENCE: Level III.


Assuntos
Artroplastia do Ombro/efeitos adversos , Artroscopia/métodos , Infecções Relacionadas à Prótese/diagnóstico , Prótese de Ombro/efeitos adversos , Adulto , Idoso , Biópsia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Propionibacterium acnes/isolamento & purificação , Infecções Relacionadas à Prótese/etiologia , Infecções Relacionadas à Prótese/microbiologia , Infecções Relacionadas à Prótese/patologia , Reoperação , Estudos Retrospectivos , Sensibilidade e Especificidade , Articulação do Ombro/microbiologia , Articulação do Ombro/patologia , Articulação do Ombro/cirurgia , Membrana Sinovial/microbiologia , Membrana Sinovial/patologia , Sinovite/microbiologia
10.
J Pak Med Assoc ; 69(Suppl 1)(1): S101-S107, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30697031

RESUMO

Tuberculosis continues to be one of the most challenging health problems more prevalent in developing countries. Pakistan ranks 5th in tuberculosis prevalence among the high-burden countries. Prosthetic joint infection of the knee by acid fast bacilli is a rare and distressing complication, occurring in nearly 1% of primary joint arthroplasties requiring prolonged medical treatment and multiple surgeries. A recent publication extensively reviewed English literature from 1952 to 2016, and repor ted only 64 prosthetic joint infec tion with tuberculosis, of which 27 cases involved the knee. Tuberculosis is a global health problem adding to the challenges that arthroplasty surgeons face in our resource-constrained setting. Furthermore, it presents as other inflammatory arthritis with almost same laboratory and radiological findings. The current paper was planned to highlight the preoperative and postoperative challenges that the arthroplasty surgeon may have in diagnosis and management of this rare infection. We included studies from 1996 to date which reported knee tuberculosis prosthetic joint infection that were managed by medication alone or with surgical intervention in patients who had undergone arthroplasty.


Assuntos
Antituberculosos/uso terapêutico , Artrite Infecciosa/terapia , Artroplastia do Joelho , Desbridamento , Remoção de Dispositivo , Doenças Endêmicas , Infecções Relacionadas à Prótese/terapia , Membrana Sinovial/patologia , Tuberculose Osteoarticular/terapia , Artrite Infecciosa/diagnóstico , Artrodese , Técnicas Histológicas , Humanos , Técnicas de Amplificação de Ácido Nucleico , Paquistão , Infecções Relacionadas à Prótese/diagnóstico , Reoperação , Membrana Sinovial/microbiologia , Teste Tuberculínico , Tuberculose Osteoarticular/diagnóstico
11.
J Int Med Res ; 47(2): 635-640, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30392431

RESUMO

OBJECTIVE: Osteoarthritis (OA) is a common cause of disability affecting millions of people of all ages worldwide. The pathogenesis involves an inflammatory component, but the cause of the inflammation remains incompletely understood. The intracellular bacteria Chlamydia trachomatis and C. pneumoniae have been demonstrated in patients with reactive arthritis. Both of these microorganisms can cause chronic and persistent infections, with C. trachomatis being the most common cause of reactive arthritis. This study was performed to investigate the presence of C. pneumoniae in a large number of patients with primary OA. METHODS: The study included 75 patients who underwent total knee arthroplasty. During surgery, a synovial biopsy was performed and synovial fluid drawn. Real-time polymerase chain reaction (PCR) of C. pneumoniae was run on all patients, and real-time PCR of bacterial 16S rDNA was conducted on 30 of the 75 patients to screen for the presence of other bacteria. RESULTS: Real-time PCR showed no evidence of the presence of C. pneumoniae in the patients' specimens, nor were other bacteria detected. CONCLUSIONS: Although an inflammatory component is part of the pathogenesis of OA, we found no evidence indicating that C. pneumoniae is a stimulator of that inflammation.


Assuntos
Infecções por Chlamydophila/complicações , Chlamydophila pneumoniae/isolamento & purificação , Osteoartrite/microbiologia , Líquido Sinovial/microbiologia , Membrana Sinovial/microbiologia , Sinovite/diagnóstico , Idoso , Artroplastia , Infecções por Chlamydophila/microbiologia , DNA Bacteriano/genética , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite/genética , Osteoartrite/cirurgia , Reação em Cadeia da Polimerase , Prognóstico , RNA Ribossômico 16S/genética , Sinovite/genética , Sinovite/microbiologia
12.
Sci Rep ; 8(1): 14305, 2018 09 24.
Artigo em Inglês | MEDLINE | ID: mdl-30250232

RESUMO

Human intestinal microbes can mediate development of arthritis - Studies indicate that certain bacterial nucleic acids may exist in synovial fluid (SF) and could be involved in arthritis, although the underlying mechanism remains unclear. To characterize potential SF bacterial nucleic acids, we used 16S rRNA gene amplicon sequencing to assess bacterial nucleic acid communities in 15 synovial tissue (ST) and 110 SF samples from 125 patients with rheumatoid arthritis (RA) and 16 ST and 42 SF samples from 58 patients with osteoarthritis (OA). Our results showed an abundant diversity of bacterial nucleic acids in these clinical samples, including presence of Porphyromonas and Bacteroides in all 183 samples. Agrobacterium, Comamonas, Kocuria, Meiothermus, and Rhodoplanes were more abundant in synovial tissues of rheumatoid arthritis (STRA). Atopobium, Phascolarctobacterium, Rhodotorula mucilaginosa, Bacteroides uniformis, Rothia, Megasphaera, Turicibacter, Leptotrichia, Haemophilus parainfluenzae, Bacteroides fragilis, Porphyromonas, and Streptococcus were more abundant in synovial tissues of osteoarthritis (STOA). Veillonella dispar, Haemophilus parainfluenzae, Prevotella copri and Treponema amylovorum were more abundant in synovial fluid of rheumatoid arthritis (SFRA), while Bacteroides caccae was more abundant in the synovial fluid of osteoarthritis (SFOA). Overall, this study confirms existence of bacterial nucleic acids in SF and ST samples of RA and OA lesions and reveals potential correlations with degree of disease.


Assuntos
Artrite Reumatoide/microbiologia , Bactérias/genética , Ácidos Nucleicos/análise , Osteoartrite/microbiologia , Líquido Sinovial/microbiologia , Membrana Sinovial/microbiologia , Adulto , Algoritmos , Citocinas/metabolismo , Análise Discriminante , Humanos , Microbiota , Análise de Componente Principal
13.
J Am Acad Orthop Surg ; 26(20): e442-e447, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30102651

RESUMO

INTRODUCTION: Identification of microorganisms is critical for correct management of an infected arthroplasty. Our hypothesis is that the culture yield depends on the location around the prosthesis from which samples are obtained. METHODS: This prospective study included 298 revisions of the hip (123) and knee (175). We compared the yield of the intraoperative samples obtained, which included synovial fluid (two), neosynovium (two), and periprosthetic membrane (two). RESULTS: Cultures were positive in 28 cases, in which 15 had the same diagnosis considering either the neosynovium or the membrane, and there were 3 cases in which the infection could have been diagnosed only by considering the combination of both. In all, there were 8 cases in which the infection might have been misdiagnosed unless considering a combination of both solid tissue samples (P = 0.004). CONCLUSIONS: The yields of the periprosthetic membrane and neosynovium do not differ significantly, and we recommend considering a combination of both. LEVEL OF EVIDENCE: Diagnostic Level II.


Assuntos
Artroplastia de Quadril/efeitos adversos , Artroplastia do Joelho/efeitos adversos , Infecções Bacterianas/diagnóstico , Técnicas Bacteriológicas , Infecções Relacionadas à Prótese/diagnóstico , Líquido Sinovial/microbiologia , Membrana Sinovial/microbiologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Estudos Prospectivos , Reoperação
14.
Stem Cell Res Ther ; 9(1): 80, 2018 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-29587847

RESUMO

BACKGROUND: Latent microorganism infection is a safety concern for the clinical application of mesenchymal stem cells (MSCs). The aim of this study is to investigate the frequencies and sensitivities of the latent virus and mycoplasma infections in synovium, bone marrow, peripheral blood cells, and blood plasma and cultured synovial MSCs. METHODS: Total DNA and RNA of the synovium (n = 124), bone marrow (n = 123), peripheral blood cells (n = 121), plasma (n = 121), and 14-day cultured synovial MSCs (n = 63) were collected from patients who underwent total knee arthroplasty or anterior ligament reconstruction after written informed consents were obtained. The multiplex polymerase chain reaction (PCR) primers were designed to quantitatively measure the representative genomes of 13 DNA viruses, 6 RNA viruses, and 9 mycoplasmas. Multi-spliced mRNA detection and virus spike test were also performed to demonstrate the sensitivity of synovial MSCs to the candidate pathogens. RESULTS: In synovium and bone marrow, the positive rates of parvovirus B19 genome were significantly higher than in peripheral blood cells (18.7% and 22% vs. 0.8%, respectively). Multi-alignment analysis of amplified and sequenced viral target genes showed the proximity of the parvovirus B19 gene from different tissue in the same patients. Synovial MSCs cultured for 14 days were positive for virus infection only in two patients (2/62 = 3%). Parvovirus B19 multi-spliced mRNAs were not detected in these two samples. Virus spike test demonstrated the sensitivity of synovial MSCs to herpes simplex virus (HSV)1 and cytomegalovirus (CMV), but not to parvovirus B19. CONCLUSION: This study revealed a relatively high incidence of latent parvovirus B19 in synovium and bone marrow tissue.


Assuntos
Medula Óssea/virologia , Mycoplasma/patogenicidade , Infecções por Parvoviridae/epidemiologia , Parvovirus B19 Humano/patogenicidade , Membrana Sinovial/virologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Medula Óssea/microbiologia , Células Cultivadas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mycoplasma/genética , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/epidemiologia , Infecções por Parvoviridae/diagnóstico , Parvovirus B19 Humano/genética , Membrana Sinovial/microbiologia
15.
Rheumatol Int ; 38(6): 1009-1016, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29404675

RESUMO

Enterobacterial components in the joints of patients are believed to contribute to a perpetuating inflammation leading to a reactive arthritis (ReA), a condition in which microbial agents cannot be recovered from the joint. At present, it is unclear whether nucleic acids from Shigella spp. are playing a pathogenic role in causing not only ReA but also other forms of arthritis. Quantitative real-time polymerase chain reaction assay (qPCR) is the method of choice for the identification of bacteria within the synovium. The aim of our study was to detect the presence of Shigella spp. nucleic acids in the synovial tissue (ST) of Tunisian arthritis patients. We investigated 57 ST samples from rheumatoid arthritis (RA) n = 38, undifferentiated oligoarthritis (UOA) n = 12, and spondyloarthritis (SpA) n = 7 patients; 5 ST samples from healthy individuals were used as controls. Shigella spp. DNA and mRNA transcripts encoding the virulence gene A (VirA) were examined using an optimized qPCR with newly designed primers and probes. Using qPCR, Shigella spp. DNA was found in 37/57 (65%) ST samples (24/38, i.e., 63.2% of RA, 8/12, i.e., 67% of UOA, and 5/7, i.e., 71.4% of SpA patients). Paired DNA and mRNA were extracted from 39 ST samples, whose VirA cDNA was found in 29/39 (74.4%) patients. qPCR did not yield any nucleic acids in the five healthy control ST samples. The qPCR assay was sensitive and showed a good intra- and inter-run reproducibility. These preliminary findings generated by an optimized, highly sensitive PCR assay underline a potential role of past gastrointestinal infections. In Tunisian patients, a bacterial etiology involving Shigella spp. in the manifestation of arthritic disorders including RA might be more common than expected.


Assuntos
Artrite Reumatoide/microbiologia , DNA Bacteriano/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Shigella/isolamento & purificação , Membrana Sinovial/microbiologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ácidos Nucleicos , Proibitinas , Reprodutibilidade dos Testes , Tunísia
16.
Microb Pathog ; 113: 248-264, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29074430

RESUMO

Intravenous inoculation of Swiss mice with S. aureus leads to severe synovial joint tissue swelling along with prominent T lymphocyte infiltrate with associated inflammation in synovial tissue. Cytokines released from macrophages such as TNF-α, IL-1ß and IL-6 the main players that precede cartilage and bone destruction during septic arthritis (SA) followed by osteoclast differentiation and bone resorption. CD4+ naïve T cells upon cytokine driven activation, differentiate into lineages of helper (Th) and regulatory T cells (Treg) including inflammatory Th17 cell lineage. Acting as counterbalance, Tregs protect the host by releasing anti-inflammatory IL-10. A disturbed balance between Th17 and Treg cell development skews the pathways towards Th17 lineage, but how it actually induces SA is still unexplored. Therefore, this study has been attempted to demonstrate the Th17/Treg ratio in synovial tissue, spleen and peripheral blood by FACS and their derived cytokines from serum of arthritic mice. Here, we reported that the ratios of Th17/Treg as well as their related cytokine levels were increased at 3 days post-infection which was decreased during 9 DPI but heightened again at 15DPI resulting in persistence of the disease, though decreased again at 30 DPI even in animals with increased dose of infection. Bacterial colonies were present in synovial joints at 15 DPI in animals with increased infection but found to be absent at 30 DPI. Maintaining Th17/Treg balance by neutralizing functionally active Th17 and their related cytokines or adoptive transfer of fully active Tregs and/or their related cytokines may lead to a novel therapeutic strategy for combating Staphylococcal arthritis.


Assuntos
Artrite Infecciosa/imunologia , Artrite Infecciosa/microbiologia , Citocinas/metabolismo , Staphylococcus aureus/patogenicidade , Linfócitos T Reguladores/fisiologia , Células Th17/fisiologia , Animais , Artrite Infecciosa/sangue , Artrite Infecciosa/fisiopatologia , Reabsorção Óssea , Linfócitos T CD4-Positivos , Diferenciação Celular , Contagem de Colônia Microbiana , Modelos Animais de Doenças , Inflamação , Interleucina-10/metabolismo , Interleucina-17/metabolismo , Interleucina-23/metabolismo , Linfócitos , Macrófagos/imunologia , Macrófagos/microbiologia , Masculino , Camundongos , Osteoclastos , Baço/microbiologia , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/microbiologia , Membrana Sinovial/microbiologia , Fator de Necrose Tumoral alfa/metabolismo
17.
Hip Int ; 27(6): 595-598, 2017 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-28731488

RESUMO

INTRODUCTION: Cultures of deep synovial biopsies remain an important tool in diagnosing periprosthetic joint infection, a devastating complication following total hip arthroplasty (THA). Recent reports of unexpected positive intraoperative cultures in aseptic revision arthroplasty, however, challenge the validity and interpretation of these cultures. The aim of this study was to evaluate the contamination risk of synovial biopsy cultures collected intraoperatively during primary THA of healthy subjects. METHODS: Synovial biopsies for culture were collected during primary total hip arthroplasty procedures from 100 consecutive cases. The synovial biopsies were taken within the first 15 minutes after skin incision. Biopsy specimen were cultured on 4 different media for 8 or 15 days. Positive cultures were identified using Maldi-Tof spectrometry. RESULTS: 16 cultures yielded a bacterium, suggesting a false positive result of 16%. The mean time for the cultures to become positive was 6.29 days (standard deviation [SD] 3.90) with a maximum of 15 days. Proprionibacterium acnes and Staphylococcus epidermidis were most commonly cultured with 6 positive results for both bacteria. CONCLUSIONS: Our study yielded a 16% false positive rate in cultures of synovial biopsy taken during primary total hip arthroplasty of healthy subjects, suggesting that contamination risk of these synovial biopsy cultures may be larger than assumed by clinicians.


Assuntos
Artroplastia de Quadril/efeitos adversos , Bactérias/isolamento & purificação , Biópsia/métodos , Prótese de Quadril/efeitos adversos , Infecções Relacionadas à Prótese/diagnóstico , Membrana Sinovial/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Infecções Relacionadas à Prótese/microbiologia
18.
Int Orthop ; 41(7): 1307-1313, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28160020

RESUMO

PURPOSE: Diagnosing periprosthetic infection remains a challenge. Multiplex-PCR and biomarkers such as alpha-defensin are potentially useful and fast methods for detecting periprosthetic infection. This study compared these new methods with clinical assessment, conventional microbiological methods and histo-pathological examination. METHODS: Twenty-eight consecutive patients with 30 joints and a mean age of 67.7 years (range 39 to 88) with removal of total hip arthroplasty (THA) or total knee replacement (TKR) were included in this study. Patients were classified according to the modified Musculoskeletal Infection Society score (MSIS) for infected joints. Punction fluid and tissue specimens were taken for conventional microbiological examination, alphadefensin test was performed, a synovial membrane specimen was used for multiplex-PCR and histopathological examination was carried out. RESULTS: The alpha-defensin test and multiplex-PCR showed a sensitivity of 76.9 vs. 30.8% and a specificity of 82.4 vs. 100%, respectively. We found a significant difference between the positive and negative results (p = 0.0023). The conventional microbiological methods were not significantly different from the alpha-defensin test (p = 0.244) with a sensitivity of 84.6% and a specificity of 100% but did differ significantly from the multiplex PCR (p = 0.0030). There was a significant difference between modified MSIS classification and multiplex PCR (p = 0.0007). CONCLUSIONS: Neither alpha-defensin test nor multiplex-PCR could detect periprosthetic infection immediately and reliably. Multiplex-PCR was suitable for detecting the non-infected but not the truly infected. Alpha-defensin test was helpful but showed no satisfactory results. Conventional microbiological methods remain the most reliable for periprosthetic infection diagnosis.


Assuntos
Artroplastia de Quadril/efeitos adversos , Artroplastia do Joelho/efeitos adversos , Reação em Cadeia da Polimerase/métodos , Infecções Relacionadas à Prótese/diagnóstico , Membrana Sinovial/microbiologia , alfa-Defensinas/análise , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/análise , Remoção de Dispositivo , Feminino , Humanos , Prótese Articular/microbiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Reprodutibilidade dos Testes , Líquido Sinovial/química
19.
Int Orthop ; 41(7): 1355-1359, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28091766

RESUMO

BACKGROUND: Histopathological tissue analysis is a key parameter within the diagnostic algorithm for suspected periprosthetic joint infections (PJIs), conventionally acquired in open surgery. In 2014, Hügle and co-workers introduced novel retrograde forceps for retrograde synovial biopsy with simultaneous fluid aspiration of the knee joint. We hypothesised that tissue samples acquired by retrograde synovial biopsy are equal to intra-operatively acquired deep representative tissue samples regarding bacterial detection and differentiation of periprosthetic infectious membranes. METHOD: Thirty patients (male n = 15, 50%; female n = 15, 50%) with 30 suspected PJIs in painful total hip arthroplasties (THAs) were included in this prospective, controlled, non-blinded trial. The results were compared with intra-operatively obtained representative deep tissue samples. RESULTS: In summary, 27 out of 30 patients were diagnosed correctly as infected (17/17) or non-infected (10/13). The sensitivity to predict a PJI using the Retroforce® sampling forceps in addition to standard diagnostics was 85%, the specificity 100%. CONCLUSIONS: Retrograde synovial biopsy is a new and rapid diagnostic procedure under local anaesthesia in patients with painful THAs with similar histological results compared to deep tissue sampling.


Assuntos
Artroplastia de Quadril/efeitos adversos , Biópsia/métodos , Estudos Prospectivos , Infecções Relacionadas à Prótese/diagnóstico , Membrana Sinovial/microbiologia , Idoso , Biópsia/instrumentação , Feminino , Humanos , Masculino , Cuidados Pré-Operatórios , Infecções Relacionadas à Prótese/microbiologia , Infecções Relacionadas à Prótese/cirurgia , Instrumentos Cirúrgicos , Líquido Sinovial/microbiologia , Membrana Sinovial/patologia
20.
Expert Rev Clin Immunol ; 13(3): 197-206, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27627462

RESUMO

INTRODUCTION: Reactive arthritis (ReA) is an inflammatory disease that can follow gastrointestinal or genitourinary infections. The primary etiologic agent for post-venereal ReA is the bacterium Chlamydia trachomatis; its relative, C pneumoniae, has also been implicated in disease induction although to a lesser degree. Studies have indicated that the arthritis is elicited by chlamydiae infecting synovial tissue in an unusual biologic state designated persistence. We review clinical aspects, host-pathogen interactions, and treatments for the disease. Areas covered: We briefly discuss both the historic and,more extensively, the current medical literature describing ReA, and we provide a discussion of the biology of the chlamydiae as it relates to elicitation of the disease. A summary of clinical aspects of Chlamydia-induced ReA is included to give context for approaches to treatment of the arthritis. Expert commentary: Basic research into the biology and host-pathogen interactions characteristic of C trachomatis has provided a wealth of information that underlies our current understanding of the pathogenic processes occurring in the ReA synovium. Importantly, a promising approach to cure of the disease is at hand. However, both basic and clinical research into Chlamydia-induced ReA has lagged over the last 5 years, including required studies relating to cure of the disease.


Assuntos
Artrite Infecciosa/diagnóstico , Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/imunologia , Chlamydophila pneumoniae/imunologia , Membrana Sinovial/imunologia , Animais , Antibacterianos/uso terapêutico , Artrite Infecciosa/terapia , Infecções por Chlamydia/terapia , Quimioterapia Combinada , Interações Hospedeiro-Patógeno , Humanos , Proibitinas , Membrana Sinovial/microbiologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA