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Abstract

OBJECTIVE

To validate the use of a flow cytometric assay that uses 2‘,7‘-dichlorodihydrofluorescein diacetate (DCFH-DA) to measure reactive oxygen species in the erythrocytes of healthy dogs.

ANIMALS

50 healthy adult dogs.

PROCEDURES

Erythrocytes were incubated with DCFH-DA or a vehicle control (dimethyl sulfoxide), then incubated with (stimulated) or without (unstimulated) hydrogen peroxide. The flow cytometric assay was evaluated for specificity with increasing concentrations of DCFH-DA and hydrogen peroxide, and a polynomial regression line was applied to determine optimal concentrations. For precision, samples were analyzed 5 consecutive times for determination of intra- and interassay variability. Stability of samples stored at 4°C for up to 48 hours after blood collection was determined with flow cytometric analysis. Coefficient of variation (CV) was considered acceptable at 20%. Baseline measurements were used to determine an expected range of median fluorescence intensity for unstimulated erythrocytes incubated with DCFH-DA.

RESULTS

Erythrocytes were successfully isolated, and stimulated samples demonstrated higher median fluorescence intensity, compared with unstimulated samples. The intra-assay CV was 11.9% and 8.9% and interassay CV was 11.9% and 9.1% for unstimulated and stimulated samples, respectively. Unstimulated samples were stable for up to 24 hours, whereas stimulated samples were stable for up to 48 hours.

CONCLUSIONS AND CLINICAL RELEVANCE

Flow cytometry for the measurement of reactive oxygen species in the erythrocytes of healthy dogs by use of DCFH-DA had acceptable specificity, precision, and stability. Flow cytometry is a promising technique for evaluating intraerythrocytic oxidative stress for healthy dogs.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate the effect of knot location on the biomechanical strength and gapping characteristics of ex vivo canine gastrocnemius tenorrhaphy constructs.

SAMPLE

36 cadaveric gastrocnemius tendons from 18 adult dogs.

PROCEDURES

Tendons were randomly assigned to 3 groups (12 tendons/group) and sharply transected and repaired by means of a core locking-loop suture with the knot at 1 of 3 locations (exposed on the external surface of the tendon, buried just underneath the external surface of the tendon, or buried internally between the apposed tendon ends). All repairs were performed with size-0 polypropylene suture. All constructs underwent a single load-to-failure test. Yield, failure, and peak forces, mode of failure, and forces required for 1- and 3-mm gap formation were compared among the 3 knot-location groups.

RESULTS

Mean yield, failure, and peak forces and mean forces required for 1- and 3-mm gap formation did not differ significantly among the 3 groups. The mode of failure also did not differ significantly among the 3 groups, and the majority (33/36 [92%]) of constructs failed owing to the suture pulling through the tendinous substance.

CONCLUSIONS AND CLINICAL RELEVANCE

Final knot location did not significantly affect the biomechanical strength and gapping characteristics of canine gastrocnemius tenorrhaphy constructs. Therefore, all 3 evaluated knot locations may be acceptable for tendon repair in dogs. In vivo studies are necessary to further elucidate the effect of knot location in suture patterns commonly used for tenorrhaphy on tendinous healing and collagenous remodeling at the repair site.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To determine the effect of epitendinous suture (ES) caliber on the tensile strength of flexor tendon repairs in cadaveric specimens from dogs.

SAMPLE

60 cadaveric superficial digital flexor tendons (SDFTs) from 30 skeletally mature dogs.

PROCEDURES

Specimens were randomly assigned to 5 suture caliber groups (n = 12 SDFTs/group). After sharp transection, SDFTs were repaired by placement of a simple continuous circumferential ES created with size-0, 2-0, 3-0, 4-0, or 5-0 polypropylene suture. Constructs were preloaded to 2 N and load tested to failure. Loads at yield, peak, and failure and mode of failure were compared among groups by statistical methods.

RESULTS

Yield, peak, and failure loads for SDFT repair constructs were positively correlated with ES caliber and did not differ between the size-0 and 2-0 groups on pairwise comparisons. Yield load was significantly greater for size-0, 2-0, and 3-0 groups than for the 4-0 and 5-0 groups. Peak and failure loads were significantly greater for the size-0 and 2-0 groups than for the remaining groups. Most size-0 (12/12), 2-0 (12/12), and 3-0 (10/12) group constructs failed because of ES pull-through; several constructs in the 4-0 group (5/12) and most in the 5-0 group (11/12) failed because of ES breakage.

CONCLUSIONS AND CLINICAL RELEVANCE

Results suggested size-0 and 2-0 sutures should be considered when placing an ES for flexor tendon repairs in dogs. However, in vivo studies are needed determine the effects of increasing ES caliber on clinical outcomes for dogs undergoing these procedures.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To compare the biomechanical strength and incidence of gap formation among canine superficial digital flexor tendon (SDFT) constructs that underwent core tenorrhaphy only and those in which the core tenorrhaphy was augmented with skin staples or a continuous Silfverskiold cross-stitch (SXS) suture pattern.

SAMPLE

42 cadaveric forelimb SDFTs from 21 musculoskeletally normal dogs.

PROCEDURES

Tendons were randomly assigned to 3 groups (14 SDTFs/group), sharply transected, and repaired with a core locking-loop suture alone (group 1) or augmented with circumferential placement of skin staples (group 2) or a continuous SXS suture pattern (group 3) in the epitenon. All constructs underwent a single load-to-failure test. Yield, peak, and failure loads, incidence of gap formation, and mode of failure were compared among the 3 groups.

RESULTS

Mean yield, peak, and failure loads differed significantly among experimental groups and were greatest for group 3 and lowest for group 1 constructs. The incidence of gap formation differed among the tested groups and was lowest for group 3 and highest for group 1. The most common mode of construct failure was the suture pulling through the tendon for group 1, staple deformation for group 2, and epitendinous suture breakage for group 3.

CONCLUSIONS AND CLINICAL RELEVANCE

Results indicated epitendinous placement of skin staples around a core SDFT tenorrhaphy site improved the biomechanical strength and resistance to gap formation for the repair but was inferior to epitendinous placement of SXS sutures. Further research is necessary before skin staples are used for tenorrhaphy augmentation in clinical patients.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate the effect of suture caliber on the tensile strength of tenorrhaphies performed with a locking-loop technique in cadaveric canine tendons

SAMPLE

60 superficial digital flexor tendons (SDFTs) from 30 cadaveric adult dogs.

PROCEDURES

Transverse tenotomy was performed, and SDFTs were repaired with a locking-loop technique and polypropylene suture of 5 randomly assigned calibers: size-0, 2-0, 3-0, 4-0, or 5-0 (n = 12 SDFTs/suture caliber). Tendon constructs were tested to failure. Yield, peak, and failure forces and causes of failure were compared between groups.

RESULTS

Mean ± SD failure force for the constructs was significantly greater with large-caliber suture (size-0: 73.5 ± 3.1 N; size 2-0: 54.4 ± 7.1 N; size 3-0: 28.7 ± 4.9 N; size 4-0: 18.7 ± 3.4 N; and size 5-0: 8.8 ± 2.8 N). The likelihood of construct failure by suture pullout through the tendon substance increased with large-caliber suture (size-0: 12/12), whereas the likelihood of construct failure by suture breakage increased with small-caliber suture (2-0: 10/12; 3-0, 4-0, and 5-0: 12/12 each).

CONCLUSIONS AND CLINICAL RELEVANCE

Large-caliber suture had greater tensile strength for tenorrhaphies performed with a locking-loop technique in cadaveric canine tendons. Prior to the use of large-caliber suture in patients requiring tenorrhaphy, however, in vivo studies are required to confirm the results obtained here.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate the effects of using an internal fixation plate to augment primary 3-loop pulley (3LP) repair of canine gastrocnemius tendons (GTs).

SAMPLE

48 cadaveric GTs from 24 adult dogs.

PROCEDURES

GTs were dissected free from other tissues, transected, and randomly assigned to 4 groups (n = 12/group). GTs were repaired with 2-0 polypropylene with a 3LP repair alone or a 3LP repair augmented with a 3-hole veterinary cuttable plate (3VCP), a 5-hole veterinary cuttable plate (5VCP), or a 7-hole veterinary cuttable plate (7VC P). Biomechanical loads, construct stiffness, gap formation, and failure modes were compared between groups.

RESULTS

Yield, peak, and failure loads were all significantly increased for the 5VCP and 7VCP groups, compared with the 3LP alone group. Increasing plate length from 3VCP to 5VCP and from 3VCP to 7VCP increased yield, peak, and failure loads. No differences were found between the 3LP and 3VCP groups with regard to yield and peak loads, but failure load was increased in the 3VCP group. Loads to create 1-mm and 3-mm gaps were significantly greater for the 5VCP and 7VCP groups, compared with the 3LP alone and 3VCP groups. Mode of plate attachment failure differed among groups.

CLINICAL RELEVANCE

Tendon plate augmentation may be a viable surgical option to increase the strength of the tenorrhaphy in dogs. However, in vivo studies evaluating the effects of plate augmentation on the tendon blood supply and progression of healing are needed prior to clinical application.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To determine effects of oral administration of metronidazole or doxycycline on olfactory function in explosives detection (ED) dogs.

ANIMALS 18 ED dogs.

PROCEDURES Metronidazole was administered (25 mg/kg, PO, q 12 h for 10 days); the day prior to drug administration was designated day 0. Odor detection threshold was measured with a standard scent wheel and 3 explosives (ammonium nitrate, trinitrotoluene, and smokeless powder; weight, 1 to 500 mg) on days 0, 5, and 10. Lowest repeatable weight detected was recorded as the detection threshold. There was a 10-day washout period, and doxycycline was administered (5 mg/kg, PO, q 12 h for 10 days) and the testing protocol repeated. Degradation changes in the detection threshold for dogs were assessed.

RESULTS Metronidazole administration resulted in degradation of the detection threshold for 2 of 3 explosives (ammonium nitrate and trinitrotoluene). Nine of 18 dogs had a degradation of performance in response to 1 or more explosives (5 dogs had degradation on day 5 or 10 and 4 dogs had degradation on both days 5 and 10). There was no significant degradation during doxycycline administration.

CONCLUSIONS AND CLINICAL RELEVANCE Degradation in the ability to detect odors of explosives during metronidazole administration at 25 mg/kg, PO, every 12 hours, indicated a potential risk for use of this drug in ED dogs. Additional studies will be needed to determine whether lower doses would have the same effect. Doxycycline administered at the tested dose appeared to be safe for use in ED dogs.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To compare the bone temperature and final hole dimensions associated with sequential overdrilling (SO) and single 6.2-mm drill bit (S6.2DB) methods used to create transcortical holes in the third metacarpal bones (MCIIIs) of horse cadavers.

Sample—60 MCIIIs from 30 horse cadavers.

Procedures—In phase 1, hole diameter, tap insertion torque, peak bone temperature, and postdrilling bit temperature for 6.2-mm-diameter holes drilled in the lateral or medial cortical region of 12 MCIIIs via each of three 2-bit SO methods with a single pilot hole (diameter, 3.2, 4.5, or 5.5 mm) and the S6.2DB method were compared. In phase 2, 6.2-mm-diameter transcortical holes were drilled via a 2-bit SO method (selected from phase 1), a 4-bit SO method, or a S6.2DB method at 1 of 3 locations in 48 MCIIIs; peak bone temperature during drilling, drill bit temperature immediately following drilling, and total drilling time were recorded for comparison.

Results—Hole diameter or tap insertion torque did not differ among phase 1 groups. Mean ± SD maximum bone temperature increases at the cis and trans cortices were significantly less for the 4-bit SO method (3.64 ± 2.01°C and 8.58 ± 3.82°C, respectively), compared with the S6.2DB method (12.00 ± 7.07°C and 13.19 ± 7.41°C, respectively). Mean drilling time was significantly longer (142.9 ± 37.8 seconds) for the 4-bit SO method, compared with the S6.2DB method (49.7 ± 24.3 seconds).

Conclusions and Clinical Relevance—Compared with a S6.2DB method, use of a 4-bit SO method to drill transcortical holes in cadaveric equine MCIIIs resulted in smaller bone temperature increases without affecting hole accuracy.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine the extent to which a hydroxyapatite coating promotes pin stability in the third metacarpal bone during transfixation casting in horses.

Animals—14 adult horses.

Procedures—7 horses each were assigned to either an uncoated or hydroxyapatite-coated pin group. Three transcortical pins were placed in the third metacarpal bone of each horse and incorporated into a cast for 8 weeks. Insertion and extraction torque were measured, and torque reduction was calculated. Radiography was performed at 0, 4, and 8 weeks. Lameness evaluation was performed at 2, 4, 6, and 8 weeks. Bacteriologic culture of pins and pin holes was performed at pin removal.

Results—All horses used casts without major complication throughout the study. Insertion torque was higher in uncoated pins. There was no effect of group on extraction torque. Hydroxyapatite-coated pins had lower torque reduction. Five of 15 hydroxyapatite-coated pins maintained or increased stability, whereas all uncoated pins loosened. Pin hole radiolucency, lameness grades, and positive bacteriologic culture rates were not different between groups.

Conclusions and Clinical Relevance—Hydroxyapatite coating increased pin stability within the third metacarpal bone of horses during 8 weeks of transfixation casting but did not improve pin performance on clinical assessments. Clinical use of hydroxyapatite-coated transfixation pins may result in greater pin stability; however, further research is necessary to improve the consistency of pin osteointegration and elucidate whether clinical benefits will ultimately result from this approach in horses.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine whether 14-day topical ocular administration of high doses of feline recombinant interferon omega (FelFN) or human recombinant interferon alpha-2b (HulFN) solution improves clinical disease and decreases virus shedding in cats with naturally acquired viral keratoconjunctivitis.

Animals—36 cats with upper respiratory tract disease and ocular involvement.

Procedures—Cats received 1 drop of FelFN solution (1 × 106 U/mL), HulFN solution (1 × 106 U/mL), or saline (0.9% NaCl) solution (12 cats/group) in each eye twice daily for 14 days (beginning day 1). Oropharyngeal and conjunctival swab samples were collected from each cat before (day 0) and on day 14 of treatment for virus isolation (VI) and real-time quantitative PCR (RT-qPCR) testing to detect feline herpesvirus-1 and feline calicivirus. Subjective clinical scores were recorded on days 0, 3, 7, 10, and 14.

Results—The number of cats for which feline herpesvirus-1 was detected via VI or RT-qPCR assay was generally (albeit not always significantly) lower on day 14, compared with day 0 findings; however, findings on days 0 or 14 did not differ among groups. The number of cats for which feline calicivirus was detected via VI or RT-qPCR assay did not differ significantly between days 0 and 14 for any group. Clinical scores significantly decreased over the 14-day period but did not differ among groups.

Conclusions and Clinical Relevance—In cats with naturally occurring viral keratoconjunctivitis, bilateral ocular administration of high doses of FelFN or HulFN twice daily for 14 days did not improve clinical disease or virus shedding, compared with treatment with saline solution.

Full access
in American Journal of Veterinary Research