Fentanyl and methadone used as adjuncts to bupivacaine for lumbosacral epidural analgesia in sheep.

Six healthy, female, mixed-breed 18-24-month-old sheep weighing 30-48 kg were submitted to lumbosacral epidural bupivacaine in combination with either methadone or fentanyl. Epidural catheters were placed in six sheep that were given three treatments: (Bup) bupivacaine (0.5 mg/kg) alone; (BupMet) bupivacaine (0.25 mg/kg) plus methadone (0.3 mg/kg); and (BupFent) bupivacaine (0.25 mg/kg) plus fentanyl (0.002 mg/kg). Haemodynamic variables, respiratory rate, rectal temperature, analgesia by applying a standard painful stimulus, motor block and sedative scores were compared among the three treatments. These parameters were determined before epidural administration and at 5, 10, 20, 30, 60, 90, 120 minutes after treatment administration, and then every 60 minutes thereafter until the end of analgesic effect. Parametrical data were analysed by proc glimmix (SAS) for repeated measures on time and means tested by ls-means. Non-parametrical data were analysed by Fisher's exact test. Duration of analgesia was longer with BupMet (240 minutes) compared with BupFent (180 minutes; P=0.0127), but BupMet was similar to Bup (240 minutes). Both treatments with opioids produced moderate motor blockade. BupMet and BupFent produced mild sedation. Only treatment with bupivacaine alone induced cardiovascular and respiratory rate changes that stayed within acceptable limits.

Effects of intra-articular injection of mesenchymal stem cells associated with platelet-rich plasma in a rabbit model of osteoarthritis.

The current study aims to evaluate the macroscopic and histological effects of autologous mesenchymal stem cells (MSC) and platelet-rich plasma on knee articular cartilage regeneration in an experimental model of osteoarthritis. Twenty-four rabbits were randomly divided into four groups: control group, platelet-rich plasma group, autologous MSC undifferentiated group, and autologous MSC differentiated into chondrocyte group. Collagenase solution was used to induce osteoarthritis, and treatments were applied to each group at 6 weeks following osteoarthritis induction. After 60 days of therapy, the animals were euthanized and the articular surfaces were subjected to macroscopic and histological evaluations. The adipogenic, chondrogenic, and osteogenic differentiation potentials of MSCs were evaluated. Macroscopic and histological examinations revealed improved tissue repair in the MSC-treated groups. However, no difference was found between MSC-differentiated and undifferentiated chondrocytes. We found that MSCs derived from adipose tissue and platelet-rich plasma were associated with beneficial effects in articular cartilage regeneration during experimental osteoarthritis.

Comparison of analgesic and systemic effects of bupivacaine, methadone, or bupivacaine/methadone administered epidurally in conscious sheep.

OBJECTIVE: The aim of this study was to evaluate the combination of bupivacaine and methadone administered epidurally in sheep. METHODS: Six healthy female mixed-breed sheep weighing 35-46 kg and aged 12-18 months were included. Each sheep was assigned to receive three treatments: 0.5 mg/kg 0.25% bupivacaine (BP), 0.3 mg/kg 1% methadone (MT) or 0.25 mg/kg bupivacaine and 0.15 mg/kg methadone (BPMT). All drugs were injected into the lumbosacral space through an epidural catheter. Each animal received each treatment at random. Heart rate, arterial blood pressure (systolic, diastolic and mean), respiratory rate, rectal temperature, analgesia, sedation and motor block were determined before treatment and at predetermined intervals. RESULTS: The duration of analgesia was 240, 220, and 180 min for BP, MT and BPMT, respectively (P < 0.05). Motor block for all agents was mild to moderate. None or the treatments significantly altered the heart rate, blood pressure or respiratory rate. CONCLUSION: Our findings suggest that lumbosacral epidural administration of bupivacaine, methadone or a combination of the two drugs can provide perioperative analgesia in sheep as part of their management for surgical procedures in the flank and hindlimbs.

Evaluation of pH effects on genomic integrity in adipose-derived mesenchymal stem cells using the comet assay.

The use of mesenchymal stem cells (MSCs) in experimental, clinical, and therapeutic trials has grown in recent years. However, the issue remains of whether these procedures are completely safe for transplant patients. Therefore, this study was designed and carried out with the aim of evaluating two different comet assay protocols for genomic damage pattern analysis in MSCs derived from adipose tissue. The analyzed and interpreted results suggest that genetic testing is needed to support clonal expansion safety in cell therapy procedures with MSCs. Furthermore, they also suggest that if the comet assay technique would be used as a genomic integrity screening assay, the protocol performed at pH = 12 (that yielded a frequency of damaged cells: tail intensity = 9.50 ± 0.60, tail moment = 0.0122 ± 0.0007; results are reported as means ± standard deviation) would be indicated as genomic damage, and that subsequent single-strand breaks occur at pH > 13 (frequency of damaged cells: tail intensity = 30.71 ± 4.23, tail moment = 0.0447 ± 0.0073). Our study demonstrates that, in the era of regenerative medicine, it is necessary to standardize and establish a battery of tests in order to identify genomic damage prior to MSC transplantation.

Therapeutic potential of mesenchymal stem cells to treat Achilles tendon injuries.

Rupture of the Achilles tendon diminishes quality of life. The gold-standard therapy is a surgical suture, but this presents complications, including wound formation and inflammation. These complications spurred evaluation of the therapeutic potential of mesenchymal stem cells (MSCs) from adipose tissue. New Zealand rabbits were divided into 6 groups (three treatments with two time points each) evaluated at either 14 or 28 days after surgery: cross section of the Achilles tendon (CSAT); CSAT + Suture; and CSAT + MSC. A comparison between all groups at both time points showed a statistically significant increase in capillaries and in the structural organization of collagen in the healed tendon in the CSAT + Suture and CSAT + MSC groups at the 14-day assessment. Comparison between the two time points within the same group showed a statistically significant decrease in the inflammatory process and an increase in the structural organization of collagen in the CSAT and CSAT + MSC groups. A study of the genomic integrity of the cells suggested a linear correlation between an increase of injuries and culture time. Thus, MSC transplantation is a good alternative for treatment of Achilles tendon ruptures because it may be conducted without surgery and tendon suture and, therefore, has no risk of adverse effects resulting from the surgical wound or inflammation caused by nonabsorbable sutures. Furthermore, this alternative treatment exhibits a better capacity for wound healing and maintaining the original tendon architecture, depending on the arrangement of the collagen fibers, and has important therapeutic potential.

Effects of lumbosacral epidural ketamine and lidocaine in xylazine-sedated cats.

In order to determine the analgesic and cardiovascular effects of the combination of epidural ketamine and lidocaine, 6 sedated cats were studied. Six healthy, young cats were used in a prospective randomised study. Each cat underwent 3 treatments, at least 1 week apart, via epidural injection: (1) ketamine (2.5 mg/kg), (2) lidocaine (4.0 mg/kg), and (3) ketamine (2.5 mg/kg) plus lidocaine (4.0 mg/kg). Epidural injections were administered through the lumbosacral space. Analgesia, motor block, sedation, heart rate, arterial blood pressure, respiratory rate and arterial oxygen saturation were measured. Rectal temperature was compared before and after sedation as well as after epidural administration of the drugs. Epidural administration of the ketamine/lidocaine combination induced prolonged analgesia extending from the coccygeal to the T13-L1 dermatomes, leading to severe ataxia. Cardiovascular effects were significant in all treatments: heart rate decreased, but there was a minimal reduction in arterial pressure. It was concluded that adding a dose of ketamine to epidural lidocaine in cats is feasible and effective.