Recommendations for rehabilitation after surgical treatment of cranial cruciate ligament disease in dogs: A 2017 survey of veterinary practitioners.

OBJECTIVE: To report current recommendations made by veterinarians for rehabilitation after surgical treatment of cranial cruciate ligament (CrCL) disease. STUDY DESIGN: Anonymized electronic survey. SAMPLE POPULATION: Veterinarians performing CrCL stabilization. METHODS: An electronic survey was created to collect information on general attitudes toward postoperative rehabilitation and recommendations regarding therapeutic modalities and bandaging. Quantitative data are reported by descriptive statistical analysis, percentage of responses, or mean (±SD). The recommendations for postoperative bandaging beyond 24 hours and for postoperative rehabilitation after extracapsular stabilization compared with after tibial osteotomy were tested by using Cochran-Mantel-Haenszel tests, with P < .05 considered statistically significant. RESULTS: The data analysis included 376 responses (13% response rate). Most (71%) respondents consistently recommended postoperative rehabilitation. Rehabilitation was more than twofold more likely to be recommended after extracapsular stabilization than after osteotomies (P = .0142). Most respondents did not recommend bandaging beyond 24 hours postoperatively (P = .00012). CONCLUSION: Most respondents recommended either formal or informal postoperative rehabilitation therapy. CLINICAL SIGNIFICANCE: If the survey respondents are representative of veterinarians performing CrCL surgery, the current attitude is supportive of postsurgical rehabilitation. Most respondents would welcome evidence-based guidelines for rehabilitation protocols.

Optimization of conditions for in vitro production of radical oxygen species and expression of tissue factor by canine mononuclear cells and granulocytes for use in high-throughput assays.

The purpose of this study was to optimize conditions for high throughput measurement of radical oxygen species (ROS) production and expression of tissue factor, also termed procoagulant activity, by canine leukocytes. Granulocytes and mononuclear cells were separated by density gradient centrifugation from peripheral blood collected on several occasions from three healthy large breed dogs. To determine optimal conditions for ROS production, granulocytes were incubated for 1 or 3h in PBG (PBS containing 0.5% BSA and 5mM glucose) or RPMI containing 10% fetal bovine serum (FBS); lipopolysaccharide (LPS), zymosan, peptidoglycan (PGN) and phorbol myristate acetate (PMA) were used as stimuli. ROS was assessed by conversion of the nonfluorescent dye dihydrorhodamine 123 to fluorescent rhodamine 123 by radical species released into the media. To identify optimal conditions for expression of tissue factor, mononuclear cells were incubated for 5h in RPMI containing different concentrations of heat-inactivated FBS (HI-FBS), and LPS, zymosan, PGN or PMA as stimuli. Expression of tissue factor was determined using a one-stage recalcification assay performed in an automated nephelometric coagulation analyzer. Neither LPS nor zymosan increased ROS production by granulocytes incubated in PBG media. In contrast, granulocytes incubated in RPMI had dose-dependent increases in ROS production in response to zymosan and PGN. ROS production was significantly increased by incubation with concentrations of LPS of 0.01microg/ml or greater, and by zymosan concentrations of 0.1microg/ml or greater. ROS production in response to incubation with PMA was significantly increased starting at 10(-7)M, and was significantly greater for cells incubated in RPMI than cells incubated in PBG. LPS-, zymosan- and PGN-stimulated procoagulant activity increased in a dose-dependent manner, whereas PMA-stimulated procoagulant activity peaked at 10(-7)M. Increasing concentrations of HI-FBS significantly increased LPS-, zymosan- and PGN-induced procoagulant activity of mononuclear cells. Results obtained in this study indicate production of ROS by canine granulocytes is optimal when these cells are incubated for 3h in RPMI with LPS (0.1microg/ml), zymosan (10 microg/ml), PGN (10 microg/ml), and PMA (10(-7)M). Furthermore, canine mononuclear cells express procoagulant activity in response to LPS, zymosan, PGN, and PMA, and responses to LPS, zymosan and PGN are enhanced by the addition of HI-FBS. These findings suggest that HI-FBS retains important serum proteins that facilitate interactions between each of these bacterial or yeast derived products and the mononuclear cells. Consequently, future studies regarding the regulation of procoagulant activity by canine mononuclear cells should be performed in the presence of HI-FBS. Both assays utilized in this study allow high throughput of samples, and therefore are appropriate choices for rapid screening of conditions and/or therapeutic interventions affecting the canine inflammatory system.