Effects of mesenchymal stromal cells versus serum on tendon healing in a controlled experimental trial in an equine model.

BACKGROUND: Mesenchymal stromal cells (MSC) have shown promising results in the treatment of tendinopathy in equine medicine, making this therapeutic approach seem favorable for translation to human medicine. Having demonstrated that MSC engraft within the tendon lesions after local injection in an equine model, we hypothesized that they would improve tendon healing superior to serum injection alone. METHODS: Quadrilateral tendon lesions were induced in six horses by mechanical tissue disruption combined with collagenase application 3 weeks before treatment. Adipose-derived MSC suspended in serum or serum alone were then injected intralesionally. Clinical examinations, ultrasound and magnetic resonance imaging were performed over 24 weeks. Tendon biopsies for histological assessment were taken from the hindlimbs 3 weeks after treatment. Horses were sacrificed after 24 weeks and forelimb tendons were subjected to macroscopic and histological examination as well as analysis of musculoskeletal marker expression. RESULTS: Tendons injected with MSC showed a transient increase in inflammation and lesion size, as indicated by clinical and imaging parameters between week 3 and 6 (p < 0.05). Thereafter, symptoms decreased in both groups and, except that in MSC-treated tendons, mean lesion signal intensity as seen in T2w magnetic resonance imaging and cellularity as seen in the histology (p < 0.05) were lower, no major differences could be found at week 24. CONCLUSIONS: These data suggest that MSC have influenced the inflammatory reaction in a way not described in tendinopathy studies before. However, at the endpoint of the current study, 24 weeks after treatment, no distinct improvement was observed in MSC-treated tendons compared to the serum-injected controls. Future studies are necessary to elucidate whether and under which conditions MSC are beneficial for tendon healing before translation into human medicine.

Comparison of the BACTEC MYCO/F Lytic bottle to the isolator tube, BACTEC Plus Aerobic F/bottle, and BACTEC Anaerobic Lytic/10 bottle and comparison of the BACTEC Plus Aerobic F/bottle to the Isolator tube for recovery of bacteria, mycobacteria, and fungi from blood.

The BACTEC MYCO/F Lytic blood culture bottle (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) is designed to optimize the recovery of fungi and mycobacteria; however, this bottle also supports the growth of most aerobic bacteria. We compared the MYCO/F Lytic bottle with two other BACTEC bottles and the Isolator system for the recovery of bacteria as well as fungi and mycobacteria from blood. A total of 6,108 blood culture sets were inoculated with blood obtained from adult patients. Twenty-five to 28 ml of blood collected by a phlebotomy team for each blood culture set was randomly distributed into each of four blood culture receptacles: the Isolator tube (Wampole Laboratories, Cranbury, N.J.) and three BACTEC bottles: the MYCO/F Lytic bottle, the BACTEC Plus Aerobic/F bottle, and the BACTEC Anaerobic Lytic/10 bottle. The sediment from the Isolator tube was inoculated onto chocolate agar (CA), brain heart infusion agar (BHI), and Sabouraud dextrose agar (SDA) and into a BACTEC 13A bottle. Incubation durations were as follows: MYCO/F Lytic bottle, 42 days; Plus Aerobic/F bottle, 5 days; Anaerobic Lytic/10 bottle, 5 days; sediment from Isolator tube on CA, 3 days; sediment from Isolator tube on BHI, 30 days; sediment from Isolator tube on SDA, 30 days; and sediment from Isolator tube in a BACTEC 13A bottle, 42 days. Two isolates of Histoplasma capsulatum were recovered from the Isolator tube only. Three isolates of Mycobacterium tuberculosis complex were recovered: two isolates from the MYCO/F Lytic bottle only and one isolate from the Isolator tube (whose sediment was inoculated into the BACTEC 13A bottle) only. Two isolates of Cryptococcus neoformans were recovered: one from the MYCO/F Lytic bottle only and the other from the MYCO/F Lytic bottle and the Isolator tube (whose sediment was inoculated into the BACTEC 13A bottle). For potential pathogens overall, there was a statistical difference in recovery that favored the Isolator system over the MYCO/F Lytic bottle (P = 0.0015), including statistically significant differences for Staphylococcus aureus (P = 0.0001) and Streptococcus pneumoniae (P = 0.0313). However, there was no statistically significant difference between the two blood culture systems when detection of bloodstream infection was considered. The time to detection for all potential pathogens combined was less for the MYCO/F Lytic bottle than for the Isolator system (P = 0.0004). Overall, the potential pathogen recovery was greater for the BACTEC Plus Aerobic/F bottle than for either the Isolator system (P = 0.0003) or the MYCO/F Lytic bottle (P = 0.0001). However, the BACTEC Plus Aerobic/F bottle did not recover M. tuberculosis, H. capsulatum, or C. neoformans isolates. The combination of the Isolator system and MYCO/F Lytic bottle may be useful as a selective blood culture method to optimize the recovery of fungi and mycobacteria from blood. Compared with the manual Isolator system, the MYCO/F Lytic system has the advantage of less preanalytic processing and continuous automated monitoring of bottles for growth by the BACTEC 9240 instrument.

Comparison of Du Pont Isolator and Roche Septi-Chek for detection of fungemia.

The rapid detection of fungemia in hospitalized patients is imperative, particularly for those who are immunocompromised. Our laboratory compared the Roche Septi-Chek with the Du Pont Isolator for the recovery of fungi from blood. Of 23,586 matched pairs of blood cultures, 199 were positive. The Isolator detected 178 (89.4%) and the Septi-Chek detected 119 (59.7%) of all positive isolates. The mean recovery time for the Isolator and Septi-Chek was 2.2 and 4.9 days, respectively. The Isolator detected fungemia earlier than the Septi-Chek did and was the only culture system positive in 83% of 53 patients, whereas the Septi-Chek system yielded the same results in only 13% of the patients. The Isolator provides a more rapid and sensitive method for the recovery of fungi from blood.

Evaluation of a lysis-centrifugation system for recovery of yeasts and filamentous fungi from blood.

A lysis-centrifugation system (Isolator; E. I. du Pont de Nemours & Co., Inc., Wilmington, Del.) was compared with a biphasic brain heart infusion (BHI) medium in a prospective study of 5,125 fungal blood cultures. The Isolator recovered 90.3% of the positive cultures, compared with 63.4% recovered by the biphasic BHI medium. Overall, the detection of fungemia was increased 36.6% by the Isolator. Mean recovery times for yeasts were 2.12 and 4.90 days for the Isolator and BHI bottles, respectively. Cultures of Histoplasma capsulatum required 8.0 and 24.14 days for recovery by the Isolator and BHI bottles, respectively. The Isolator provided a more rapid and sensitive means of detecting organisms associated with fungemia.

Comparison of a radiometric method (BACTEC) and conventional culture media for recovery of mycobacteria from smear-negative specimens.

The BACTEC system and three conventional media (Middlebrook 7H10, selective Middlebrook 7H11 [S7H11], and Lowenstein-Jensen [LJ] were compared for their mean recovery times and recovery rates of mycobacteria from acid-fast, smear-negative clinical specimens. Of the 71 smear-negative, culture-positive specimens recovered from 2,165 submitted smear-negative cultures, the BACTEC system detected 71.8%, compared with 88.7% for the conventional three-medium system. When media were individually compared, BACTEC medium (Middlebrook 7H12) was more successful in recovering mycobacteria (71.8%) than was LJ (62%), Middlebrook medium 7H10 (55.9%), or Middlebrook S7H11 medium (52.1%). Middlebrook 7H11 medium containing sodium selenate was also evaluated and did not increase the recovery rate or decrease the recovery time of mycobacterial species when compared with LJ, Middlebrook 7H10, S7H11, and 7H12 media. The mean detection time for the BACTEC system was less than that by conventional methods for the seven species of mycobacteria recovered. Detection times for Mycobacterium tuberculosis on the BACTEC system and conventional cultural systems were 13.7 and 26.3 days, respectively.

Rapid urea broth test for yeasts.

A rapid, miniaturized, urea broth test useful for detecting urease activity of yeasts was compared to Christensen urea agar. All urease-producing yeasts tested were positive on both media; however, 60% were reactive in the urea R broth within 30 min, and the remainder were reactive within 4 h. This urea multiwell test may be useful as a rapid screening method for detecting urease-producing yeasts recovered from clinical specimens and as an adjunct test with other rapid methods of yeast identification.

Evaluation of a hypertonic sucrose medium for the detection of fungi in blood cultures.

A comparison was made between biphasic brain heart infusion medium without and with 15% sucrose in vented blood culture bottles. A higher recovery rate (P less than 0.01) of fungi was noted in the medium without sucrose.

Recovery of yeast from vented blood culture bottles.

Rates of isolation of yeasts from blood cultures were significantly enhanced by venting vacuum blood culture bottles in studies of both stimulated and patients' blood cultures; however, the time interval to detection of positivity of yeasts in the clinical studies was significantly (P less than 0.01) shorter in a vented bottle with biphasic brain heart infusion medium than in a vented bottle with soybean-casein digest broth. The mean time intervals to detection of positivity were 2.6 days in the former and 5.2 days in the latter.

Microbiological study of streptococcal bacteremia.

During a 2.5-year study, streptococci were isolated from 280 patients (19% of those with bacteremia). Of this number, 54 had group D streptococci (45 were enterococci), 218 had alpha- or gamma-hemolytic nongroup D streptococci, and 49 had beta-hemolytic streptococci.