Kingella kingae as the Main Cause of Septic Arthritis: Importance of Molecular Diagnosis.

BACKGROUND: Kingella kingae is an emergent pathogen causing septic arthritis (SA) in children.The objective of this study was to analyze the etiology of SA in children before and after the implementation of universal 16S rRNA gene polymerase chain reaction and sequencing (16SPCR) in synovial fluid. METHODS: Children ≤14 years with acute SA from a Madrid cohort (2002-2013) were reviewed. Differences in etiology were analyzed before (period 1) and after (period 2) the implementation of bacterial 16SPCR in 2009. A comparison in epidemiology, clinical syndromes, therapy and outcome between infections caused by K. kingae and other bacteria was performed. RESULTS: Bacteria were detected from 40/81 (49.4%) children, with a higher proportion of diagnosis after 16SPCR establishment (period 2, 63% vs. period 1, 31.4%; P = 0.005). The main etiologies were Staphylococcus aureus (37.5%) and K. kingae (35%), although K. kingae was the most common microorganism in P2 (48.3%). Children with K. kingae SA were less likely to be younger than 3 months (0 vs. 42.3%; P < 0.001), had less anemia (21.4 vs. 50%; P = 0.010), lower C-reactive protein (3.8 vs. 8.9 mg/dL; P = 0.039), less associated osteomyelitis (0 vs. 26.9%; P = 0.033), shorter intravenous therapy (6 vs. 15 days; P < 0.001), and had a nonsignificant lower rate of sequelae (0 vs. 30%; P = 0.15) than children with SA caused by other bacteria. However, they tended to have higher rate of fever (86 vs. 57%; P = 0.083). CONCLUSIONS: K. kingae was frequently recovered in children with SA after the implementation of bacterial 16SPCR, producing a milder clinical syndrome and better outcome. Therefore, the use of molecular techniques may be important for the management of these children.

Bone marrow mesenchymal stem cell aspirates from alternative sources: is the knee as good as the iliac crest?

INTRODUCTION: The most common method to obtain human mesenchymal stem cells (MSCs) is bone marrow aspiration from the iliac crest, but MSCs have also been isolated from different bones. The main purpose of this study was to compare bone marrow MSCs aspirated from the metaphysis of the distal femur and the proximal tibia with those obtained from the iliac crest, and to determine whether these locations represent potential alternative sources of MSCs for research and clinical application. MATERIALS AND METHODS: Bone marrow was aspirated from the iliac crest and the metaphysis of the distal femur and the proximal tibia during total knee arthroplasty in 20 patients. The aspirates were centrifuged by density gradient, then mononucleated cell (MNC) concentration in the different aspirates was determined using a Coulter counter. MSCs were isolated, cultivated and characterised by their immunophenotype and by their in vitro potential for differentiation into osteoblasts, chondroblasts and adipocytes in specific media. Expansion and cell viability were quantified using trypan blue staining and cell counting with a haemocytometer (Neubauer chamber). The three sources were compared in terms of MNC concentration, viability of the cultures and presence of MSC using the Wilcoxon test. RESULTS: MNC concentration was significantly higher in the iliac crest (10.05 Millions/ml) compared with the femur (0.67 Millions/ml) and tibia (1.7 Millions/ml). Culture success rates were 90%, 71% and 47% for MSCs from the iliac crest, femur and tibia, respectively. Flow cytometry analysis showed the presence of CD90+, CD105+, CD73+, VEGF+, CD71+, HLA-DR-, CD45-, CD34-, CD19-, and CD14- cells. The immunophenotype pattern of MSCs was similar for the three locations. Trilineage differentiation was achieved with all samples. CONCLUSIONS: MSCs can be found in bone marrow from the metaphysis of both the distal femur and the proximal tibia. The phenotype and differentiation potential of these cells are similar to those of bone marrow MSCs from the iliac crest. Bone marrow aspiration from these locations is a relatively easy and safe alternative to that from the iliac crest for obtaining MSCs. Further study is required to assess whether the concentrations of MSCs obtained from these sources are sufficient for one-step therapeutic purposes.