Immunohistochemical analysis of cyclooxygenase-2 induction during wound healing in dog skin

Cyclooxygenase (COX)-2 is an inducible isoform of COX and is expressed under abnormal health conditions. This study elucidated the cutaneous induction of COX-2 during the wound healing processes in dog skin. Dog skin was sutured after punch biopsy and investigated histologically and immunohistochemically on days 0 (normal), 1, 3, 5, 7, 10, and 14 after injury. Histological changes, including infiltration of inflammatory cells and proliferation of fibroblast-like cells, were observed as predicted, and there was a close and significant correlation between these 2 events. COX-2-positive cells were detected in the epidermis between days 1 and 7, and bimodal peaks were observed in the case of the percentage of COX-2-positive cells. In inflammatory cells, COX-positive signals were detected on day 3 only. Here, we clarified the localization and pattern of the induced COX-2 expression during wound healing in dog skin.     

Immunohistochemical analysis of cyclooxygenase-2 induction during wound healing in dog skin

Cyclooxygenase (COX)-2 is an inducible isoform of COX and is expressed under abnormal health conditions. This study elucidated the cutaneous induction of COX-2 during the wound healing processes in dog skin. Dog skin was sutured after punch biopsy and investigated histologically and immunohistochemically on days 0 (normal), 1, 3, 5, 7, 10, and 14 after injury. Histological changes, including infiltration of inflammatory cells and proliferation of fibroblast-like cells, were observed as predicted, and there was a close and significant correlation between these 2 events. COX-2-positive cells were detected in the epidermis between days 1 and 7, and bimodal peaks were observed in the case of the percentage of COX-2-positive cells. In inflammatory cells, COX-positive signals were detected on day 3 only. Here, we clarified the localization and pattern of the induced COX-2 expression during wound healing in dog skin.     

Effect of flunixin meglumine on surgical wound strength and healing in the rat

Forty male adolescent Sprague-Dawley rats were anesthetized and standardized ventral midline laparotomies and uniform-length gastrotomies and typhlotomies were performed. The visceral and abdominal surgically inflicted wounds were closed with 5-0 polypropylene and 4-0 nylon suture, respectively. The rats were allotted into 4 groups (10 rats/group); 2 groups were not given flunixin meglumine (controls) and 2 groups were given flunixin meglumine (1.1 mg/kg of body weight, IM, every 12 hours). On day 5 and again on day 14 after surgery, 1 control and 1 flunixin meglumine-treated group were euthanatized. Tensile strength of the skin and linea alba incisions was determined, using a computerized tensiometer. Gastric and cecal incision bursting strengths were determined, using a pressure manometer. Flunixin meglumine significantly (P less than 0.05) decreased the tensile strength of wounds in the skin and linea alba, but did not affect visceral bursting strength at day 5 after surgery. At day 14 after surgery, a significant difference in wound strength was not found between the flunixin meglumine and control groups in any of the tissues evaluated. Flunixin meglumine had an adverse influence on the inflammatory stage of wound repair, but not on the proliferative stage, when fibroplasia is a major factor in wound strength. Major histologic differences were not found in the incision wounds of flunixin meglumine-treated and nontreated control rats.     

Comparison of the role of the subcutaneous tissues in cutaneous wound healing in the dog and cat

Objective— To describe and compare the contribution of the subcutaneous tissues to 1st and 2nd intention cutaneous wound healing in the dog and cat.
Study Design— Experimental study.
Animals— Domestic shorthaired cats (n=6) and 6 beagle dogs.
Methods— Paired wounds were created on either side of the dorsal midline; the subcutaneous tissue was removed on 1 side and left intact on the other. Square, open wounds of the dorsal aspect of the thorax were observed for 21 days to monitor granulation tissue formation, wound contraction, epithelialization, and total healing (contraction+epithelialization). Breaking strength of sutured linear wounds was measured 7 days after wounding. Laser-Doppler perfusion imaging (LDPI) was used to measure cutaneous perfusion.
Results— First intention healing: subcutaneous tissue removal had no consistent effect on sutured wound strength at 7 days in dogs or cats. Second intention healing: removal of subcutaneous tissue reduced wound perfusion, granulation, contraction, epithelialization, and total healing. Granulation tissue formation and wound contraction were delayed to a significantly greater degree in cats than in dogs ( P <.05). Two dogs (33%) had minor wound infections.
Conclusions— The subcutaneous tissues make an important contribution to 2nd intention cutaneous healing. Dog and cat wounds had delayed 2nd intention healing when subcutaneous tissues were removed; wounds in dogs, but not cats, had largely recovered from this delay by 21 days.
Clinical Relevance— Extensive debridement of subcutaneous tissue may delay wound healing particularly in feline patients. A higher risk for wound infections may accompany extensive removal of subcutaneous tissues in dogs.     

Cutaneous wound healing in the cat: a macroscopic description and comparison with cutaneous wound healing in the dog

OBJECTIVE: To describe the macroscopic features of first and second intention cutaneous wound healing in the cat and compare with the dog. STUDY DESIGN: Experimental study. ANIMALS: Domestic shorthaired cats (6) and beagle dogs (6). METHODS: Square, open cutaneous wounds created on the dorsal aspect of the thorax were evaluated for 21 days for temporal and spatial development of granulation tissue, wound contraction, epithelialization, and total healing. To evaluate first intention healing, breaking strength of sutured linear cutaneous wounds was measured at 7 days post-wounding. Laser-Doppler perfusion imaging was used to measure cutaneous perfusion. RESULTS: First intention healing: sutured wounds in cats were only half as strong as those in dogs at day 7 (0.406 versus 0.818 kg breaking strength). Second intention healing: cats produced significantly less granulation tissue than dogs, with a peripheral, rather than central distribution. Wound epithelialization and total wound healing (total reduction in open wound area from contraction and epithelialization) were greater for dogs than for cats over 21 days. Wound contraction on day 7 was greater for dogs, but not on day 14 or 21. Cutaneous perfusion was initially greater for dogs than for cats, but no differences were detected after day 7. CONCLUSIONS: Significant, previously unreported differences in cutaneous wound healing exist between cats and dogs. In general, cutaneous wounds in cats are slower to heal. Cats and dogs also appear to use different mechanisms of second intention healing. In cats wounds close mainly by contraction of the wound edges, whereas in dogs wounds close more from central pull, and epithelialization. CLINICAL RELEVANCE: Surgeons should view the cat as a unique species, which presents its own special challenges in wound healing, and should take this into account when planning treatment of feline wounds, either by primary closure, or by second intention healing.     

The effects of topical tripeptide copper complex and helium-neon laser on wound healing in rabbits

The aim of this study was to compare the clinical and histopathological effects of tripeptide copper complex (TCC) and two different doses of laser application (helium-neon laser, 1 and 3 J cm(-2)) on wound healing with untreated control wounds. Experimental wounds were created on a total of 24 New Zealand white rabbits and topical TCC or laser was applied for 28 days. The wounds were observed daily, and planimetry was performed on days 7, 14, 21 and 28 to measure the unhealed wound area and percentage of total wound healing. Biopsies were taken weekly to evaluate the inflammatory response and the level of neovascularization. The median time for the first observable granulation tissue was shorter (P < 0.05) in the low and high dose laser groups than in the control group (3 and 2.66 vs. 4.5 days), but was not different between the TCC and control groups (4.16 vs. 4.5 days). Filling of the open wound to skin level with granulation tissue was faster (P < 0.05) in the TCC and high dose laser groups than in the control group (14 and 16 vs. 25 days), but was not different between the low dose laser and control groups (23 vs. 25 days). The average time for healing was shorter (P < 0.05) in the TCC and high dose laser groups (29.8 and 30.2 vs. 34.6 days), but was not different between the low dose laser and control groups (33.8 vs. 34.6 days). Histopathologically, wound healing was characterized by a decrease in the neutrophil counts and an increase in neovascularization. The TCC and high dose laser groups had greater neutrophil and vessel counts than in the control group, suggesting a more beneficial effect for wound healing.     

Electrical stimulation on skin wound healing in the horse: preliminary studies

The effect of low-level direct-current stimulation on skin wound healing in the horse was assessed. Self-sustaining electrical circuits with electrodes were implanted subcutaneously in or near the wound. Stimulation by direct current (10 or 20 microA) was used to determine the effect on equine skin healing. The efficacy of electrotherapy was evaluated by sequentially comparing the clinical appearance of the wound and measuring the size of the granulating wound bed. The histologic appearance of the healing stimulated wounds was compared with that in nonstimulated control wounds created on 9 horses. Seemingly, electrical stimulation had no discernible effect on experimentally created skin wounds. Clinical observation and histologic examination of the wounds indicated that severe tissue reaction from the implanted electrodes and concurrent local infection produced local detrimental effects to wound healing.     

Development of surgical techniques for preparation of in vitro-isolated perfused porcine skin flaps for percutaneous absorption studies

We developed a single-pedicle, axial pattern tubed skin flap that could be transferred to an in vitro perfusion apparatus. On the basis of results of prosections, angiography, contact radiography, and surviving-length studies, it was concluded that a single-pedicle, axial pattern skin flap measuring 4 cm x 12 cm incorporating the caudal superficial epigastric artery would survive to its entire length. Subsequently, a surgical (stage 1) procedure was developed for the routine preparation of single-pedicle, axial pattern tubed skin flaps. Healing after the stage-1 procedure was evaluated by visual inspection and fluorescein angiography. Stage-1 procedures were performed successfully 149 of 160 (93%) times. A second surgical (stage 2) procedure was developed for routine cannulation of the caudal superficial epigastric artery and harvest of the tubed skin flap. Stage-2 procedures were performed successfully 136 of 144 (94%) times.     

The effects of cancer and cancer therapies on wound healing

Based on experimental evidence in rodents, most of the antineoplastic agents will affect wound healing. With most of the agents, this impairment is not sufficient to produce increased morbidity based on the clinical reports in humans. Radiation therapy appears to inhibit healing in both experimental animals and during clinical trials. In spite of this, it is reported that wounds in animals will heal when they are receiving radiation therapy after surgery. Based on the information presented here and experience at the University of Missouri, the decision to use adjuvant therapy should depend on the surgery performed. With a single incision that had no increased tension, there should be no hesitation to use adjuvant therapy. If removal of the tumor required reconstructive surgery, no radiation or chemotherapy should be used until the wound has healed.     

Evaluation of the effectiveness of an ointment based on Alkannins/Shikonins on second intention wound healing in the dog

The enantiomeric naphthoquinones alkannins and shikonins (A/S) have been established as potent wound healing agents. The purpose of this study was to evaluate the effectiveness of an A/S based ointment for humans on second intention wound healing in the dog, as compared to wound flushing with Lactated Ringer's solution (LRS). Ten mixed breed dogs, aged 2 to 5 y, were used. One 2.5 × 2.5 cm full-thickness skin defect was created on the lateral aspect of each arm for subjective evaluation, laser-Doppler flowmetry (LDF), and planimetry. Additionally, 3 matching 2 × 2 cm wounds were created on opposite sides of the dorsal midline for histologic evaluation. Wounds were treated once daily with the A/S based ointment on the right side and by flushing with LRS on the left until healed (about 20 d). During the healing process, tissue perfusion (mean LDF value) was found to be significantly higher on the side treated with the A/S based ointment compared with the LRS-treated side. Histologically, angiogenesis (on days 4 and 11), collagen production score (on days 4, 11, and 20), and epithelial thickness score (on day 11) were significantly higher in the wounds treated with the A/S based ointment. Wound size, as evaluated by planimetry, decreased significantly from day 0 to day 20 on both sides, but no significant differences were found between the A/S based ointment and LRS-treated wounds.     

Comparison of the incidence of residual shunting between two surgical techniques used for ligation of patent ductus arteriosus in the dog

OBJECTIVE: To compare the incidence of residual patent ductus arteriosus (PDA) flow after ligation using 2 different dissection techniques: a standard dissection and a method described by Jackson and Henderson. STUDY DESIGN: A randomized, prospective study. ANIMALS: Thirty-five dogs admitted for surgical correction of a left to right shunting PDA. METHODS: Dogs were randomly assigned: 19 to a standard dissection technique (group S) and 16 to the Jackson and Henderson dissection group (group JH). RESULTS: Gender ratio, age at surgery, and diameter of the ductus were not statistically different between groups. Breed distribution was also similar. Because 1 dog had fatal intraoperative hemorrhage, only 34 dogs were available for residual flow comparisons. Twenty-one percent of group S dogs had residual flow compared with 53% in group JH. Whereas no intraoperative complications occurred in group S, 3 were encountered in group JH. CONCLUSIONS: The incidence of residual flow was higher when the Jackson and Henderson dissection was used for PDA ligation compared with a standard method of dissection. This was probably because of entrapment of loose connective tissue within the medial aspect of the ligature, impeding complete closure of the ductus. CLINICAL RELEVANCE: Ideal PDA closure should result in no residual ductal flow to prevent possible adverse long-term sequelae, such as recanalization and infective endocarditis.     

Comparison of conventional and non-conventional techniques for the diagnosis of bovine brucellosis in Sudan

The objective of the present study was to detect brucellosis in suspected dairy cattle in Khartoum State, Sudan using the conventional serological tests and tests done on milk in comparison to a PCR-based technique. Milk and blood samples collected simultaneously from suspected brucellosis cows (n = 147) in 12 different dairy farms around Khartoum State were used in the study. Overall, 54 (36.7%) of the total milk samples were positive according to the milk ring test (MRT), while 29 (19.7%) of the serum samples were positive according to the Rose Bengal test (RBT); microscopy on modified Ziehl-Neelsen-stained slides detected 13.6% of the cases, and recovery of Brucella species on both Brucella medium and tryptic soya agar was 7.5%. Thirty-three (22.4%) samples were found positive on PCR-amplified IS711 which were then taken as positive brucellosis cases. The differences of RBT and PCR-IS711 from MRT were highly significant (P < 0.05). MRT detected more cases of bovine brucellosis compared to RBT, PCR, microscopy, and culture. MRT is recommended as a noninvasive test compared to RBT, and it is less expensive compared to PCR and culture.     

Comparison of the healing of prescrotal urethrotomy incisions in the dog: sutured versus nonsutured

Comparison was made of the healing of sutured prescrotal urethral incisions (12 dogs) with that of nonsutured incisions (6 dogs). Comparison was also made of the healing of 5-0 polyglactin 910-sutured urethral incisions (6 dogs) with that of 5-0 polydioxanone-sutured incisions (6 dogs). Three dogs from each treatment group were euthanatized 3 weeks and 6 weeks after the surgical procedure. Surgical sites were examined grossly, and urethral circumference measurements were taken at 3 locations (surgical site, 1 cm cranially, and 1 cm caudally). Transverse sections of the surgical sites were prepared and examined by light microscopy. Hemorrhage occurred postoperatively in dogs in which the incisions were not sutured. The surgical sites from the 6 dogs in which incisions healed by second intention had more fibrosis and less inflammation than did those that were sutured. There was little difference between incisions sutured with polyglactin 910 and those sutured with polydioxanone suture material. Postoperative urethral stricture formation did not occur in any of the dogs.     

Comparison of single-dose and conventional trimethoprim-sulfadiazine therapy in experimental Staphylococcus intermedius cystitis in the female dog

Cystitis was produced in 4 groups of 6 female dogs each, using salicylic acid, ethanol, and Staphylococcus intermedius. Group-I dogs served as nontreated controls. Starting 2 days after infection was induced, group-II dogs were treated with trimethoprim-sulfadiazine at a dosage of 15 mg/kg given orally 2 times a day for 21 days; groups-III and -IV dogs were treated with single oral dosages of the antibiotic at 60 mg/kg and 90 mg/kg, respectively. Group-I dogs (controls) remained infected for the 26-day duration of the study. The response to therapy seen in group-II dogs was better than the therapeutic responses in groups-III and -IV dogs (P less than 0.05). Results of the present study do not support the efficacy of single-dose therapy for this model of cystitis.     

The effect of low level laser therapy (LLLT) on wound healing in horses.

Laser therapy is used in many countries, including South Africa, for the treatment of skin wounds. Low level galium aluminium arsenide (GaAlAs) laser was administered to full thickness skin wounds (3 x 3 cm) induced surgically on the dorsal aspect of the metacarpophalangeal joints of 6 crossbred horses in a randomised, blind, controlled study. Treated wounds that received a daily laser dosage of 2 J/cm2 were compared with nontreated control wounds on the opposite leg. There were no wound complications. Both groups of wounds were cleaned daily using tap water. Wound contraction and epithelialisation were evaluated using photoplanimetry. There were no significant differences in wound contraction or epithelialisation between the laser treated and the control wounds. It was therefore concluded that laser therapy had no clinically significant effect on second intention wound healing in this study.