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- Author or Editor: Ryland B. Edwards x
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Objective—To compare 2 methods of quantitating chondrocyte viability and to determine chondrocyte response to thermal injury over time.
Sample Population—108 stifle joints from 54 adult rats.
Procedures—Cartilage from the distal aspect of the femur was treated ex vivo with radiofrequency energy at a probe setting that would result in immediate partial-thickness chondrocyte death; untreated sections served as controls. Explants were cultured, and cell viability was compared by use of lactate dehydrogenase (LDH) histochemical staining and calcein AM and ethidium homodimer-1 confocal laser microscopy (CLM) cell viability staining. Terminal deoxynucleotidyl transferase–mediated X-dUTP nick end labeling (TUNEL) was used to detect apoptosis. All labeling studies were performed 0, 1, 3, 7, 14, and 21 days after treatment.
Results—In the treated tissues, a greater percentage of viable cells were found with CLM, compared with LDH staining. This result contrasted that of control tissues in which LDH staining indicated a greater percentage of live cells than CLM. The greatest number of TUNEL-positive chondrocytes was present at day 3, declining at later time intervals.
Conclusions and Clinical Relevance—CLM and LDH histochemistry techniques yield different absolute numbers of live and dead cells, resulting in differing percentages of live or dead cells with each technique. These differences may be related to the enzymes responsible for activation in each technique and the susceptibility of these enzymes to thermal injury. Results of TUNEL indicate that apoptosis contributes to chondrocyte death after thermal injury, with a peak signal identified 3 days after insult.
Objective—To determine the critical temperature that reduces chondrocyte viability and evaluate the ability of chondrocytes to recover after exposure to the critical temperature.
Sample Population—Cartilage explants obtained from the humeral heads of 30 sheep.
Procedures—In a randomized block design, 318 full-thickness cartilage explants were collected from 30 humeral heads of sheep and cultured for up to 14 days. On the first day of culture (day 0), explants were subjected to temperatures of 37°, 45°, 50°, 55°, 60°, or 65°C for 5 minutes by heating culture tubes in a warming block. The ability for chondrocytes to recover after exposure to the critical temperature was determined by evaluating viability at days 0, 1, 3, 7, and 14 days after heating. Images were analyzed by use of confocal laser microscopy.
Results—Analysis of images revealed a significant decrease in live cells and a significant increase in dead cells as temperature increased. Additionally, the deepest layer of cartilage had a significantly lower percentage of live cells, compared with values for the 3 most superficial layers. Chondrocytes did have some ability to recover temporarily after the initial thermal insult.
Conclusions and Clinical Relevance—A strong relationship exists between increasing temperature and cell death, with a sharp increase in chondrocyte death between 50° and 55°C. Chondrocytes in the deepest cartilage layer are most susceptible to thermal injury. The threshold of chondrocyte recovery from thermal injury is much lower than temperatures reached during chondroplasty by use of most radiofrequency energy devices.
Objective—To compare effects of synovectomy performed by use of monopolar radiofrequency energy (MRFE) versus mechanical debridement in rabbits with induced inflammatory arthritis.
Animals—25 mature female New Zealand White rabbits.
Procedure—Inflammatory arthritis was induced in both femoropatellar joints of each rabbit. Joints then were treated by mechanical debridement or MRFE treatment or served as sham-operated controls. Rabbits were euthanatized 2 weeks or 3 months after surgery. Biopsy specimens of synovium were analyzed by use of light microscopy.
Results—At 2 weeks after surgery, samples from MRFE-treated joints had fewer plasma cells and more heterophils than the other 2 groups and more lymphocytes than sham-operated controls, whereas samples from mechanically debrided joints had greater numbers of lymphocytes and heterophils than sham-operated controls. At 3 months after surgery, samples from MRFE-treated joints had fewer plasma cells than sham-operated controls, more heterophils than mechanically debrided and sham-operated controls, and more macrophages than mechanically debrided joints. There was no difference in synovial ablation, synovial proliferation, or fibrosis among the 3 groups at 2 weeks or 3 months after surgery.
Conclusions and Clinical Relevance—Analysis of results of this study documented a similar degree of synovial ablation when comparing use of MRFE to mechanical debridement. In rabbits with this method of induced inflammatory arthritis, there were no detectable benefits of MRFE or mechanical debridement on the synovium, compared with results for sham-operated control joints, at 2 weeks and 3 months after surgery for most of the synovial variables evaluated. ( Am J Vet Res 2004;65:573–577)
Objective—To evaluate the use of hydrothermal ablation of articular cartilage for arthrodesis in horses through investigation of the effects of joint lavage with physiologic saline (0.9% NaCl) solution (80°C) for various treatment times on chondrocyte viability in the articular cartilage of the metacarpophalangeal and metatarsophalangeal joints of cadaveric horse limbs.
Sample Population—7 pairs of metacarpophalangeal and 8 pairs of metatarsophalangeal joints from 8 Thoroughbreds.
Procedure—The horses were euthanatized for reasons unrelated to musculoskeletal disease. On a random basis, 1 joint of each pair underwent intra-articular lavage for 5, 10, or 15 minutes with heated saline solution (80°C); the other joint underwent sham treatment of similar duration with saline solution at 22°C (control). Cartilage samples from the distal articular surface of metacarpus III (or metatarsus III), the proximal surface of the proximal phalanx, and the lateral and medial proximal sesamoid bones were assessed for chondrocyte viability via confocal microscopy and viability staining following enzymatic digestion.
Results—Compared with the control joints, findings of both viability assays indicated that the percentage of sites containing viable chondrocytes in heat-treated joints was decreased. Treatment hazard ratios of 0.048 (confocal microscopy) and 0.2 (digestion assay) were estimated. Histologically, periarticular soft tissues had minimal detrimental effects after heat treatment.
Conclusions and Clinical Relevance—Ex vivo intraarticular lavage with saline solution at 80°C resulted in the death of almost all articular chondrocytes in the joint. This technique may be a satisfactory method for extensive cartilage ablation when performing arthrodesis by minimally invasive techniques. (Am J Vet Res 2005;66:36–42)
Objective—To evaluate long-term outcome of repair of femoral capital physeal fractures with 7.0-mm cannulated screws in juvenile bulls that weighed > 350 kg (770 lb).
Procedure—Medical records of affected cattle were identified and reviewed. Follow-up information was obtained by means of a written questionnaire, 1 to 15 years after discharge, from the veterinarians who supervised the postoperative care at bull stud facilities.
Results—Mean age and body weight at time of repair were 19 months (range, 11 to 27 months) and 513 kg (1,128.6 lb; range, 364 to 720 kg [800.8 to 1,584 lb]), respectively. Surgical repair was performed with 7.0- mm cannulated screws. A second surgery to replace malpositioned implants was required in 1 bull. Fourteen of the 20 bulls were considered serviceable for semen collection after surgical repair, and persistent lameness was evident in 11 of the 14. Mean duration for bulls to become serviceable was 5.5 months (range, 2 to 11 months). Age, weight, duration of injury at the time of repair, and degree of reduction did not have a significant effect on whether surgical repair was successful. Six bulls remained severely lame and were euthanatized.
Conclusions and Clinical Relevance—Results indicated that surgical repair of femoral capital physeal fractures in adolescent bulls with 7.0-mm cannulated screws placed in lag fashion has a good prognosis for long-term function in a semen collection facility. (J Am Vet Med Assoc 2005;227:964–969)
Objective—To describe signalment; surgical findings; short-, medium-, and long-term outcome; and recurrence rate for cattle undergoing celiotomy because of jejunal hemorrhage syndrome (JHS) and to analyze risk factors associated with outcome and recurrence.
Design—Retrospective case series.
Animals—31 dairy cattle with JHS.
Procedures—Medical records were analyzed. Follow-up information was obtained from owners of cattle surviving until discharge.
Results—18 of 31 (58%) cattle undergoing celiotomy survived to initial discharge. Fifteen (48%) and 13 (42%) were alive 6 and 12 months after discharge, respectively. All 5 deaths within 12 months after discharge were attributed to JHS recurrence. Survival time was 12 to 85 months for the 13 long-term survivors. Six of 7 that died > 12 months after celiotomy did so for reasons unrelated to JHS. Recurrence rate among short-term survivors was 7 of 18; 1 of these survived long-term. A significant proportion of affected cattle were Brown Swiss, compared with proportions for other breeds. Manual massage of the bowel to break down clots was associated with a significantly higher short-term survival rate than was en-terectomy or enterotomy. Medium- and long-term survival rate was higher in cattle referred 24 to 48 hours after onset of signs. Length of obstructing blood clots was not associated with outcome. Other factors were not significantly associated with recurrence.
Conclusions and Clinical Relevance—Survival rates were higher than those in other reports. Prompt celiotomy and resolution by use of manual massage were associated with higher survival rates. In this population, JHS recurred in 7 of 18 short-term survivors.