Effect of topical ozonated sunflower oil on second intention wound healing in turtles: a randomised experimental study.

BACKGROUND: Ozone is an antimicrobial agent that in experimental and case-control studies has been found to exert a positive effect on wound healing. Wild and pet chelonians frequently present insidious wounds exhibiting secondary infections and/or delayed healing. OBJECTIVES: Evaluate the effects of topical ozonated sunflower oil on second-intention healing of acute experimental skin wounds in red-eared sliders (Trachemys scripta elegans). METHODS: Randomised within-subject controlled study; Group 1 (n = 24) was used to assess clinical healing features; Group 2 (n = 12) was used for histological evaluation in which two sets of wounds were biopsied at 2, 7, 14, 21, 28 and 42 days over the course of the cicatrisation process. A single 6 mm diameter wound was made on each rear limb and topical ozonated (950 peroxide value) and non-ozonated sunflower oil were applied daily for one week on treated and contralateral control wounds, respectively. RESULTS: Mean wound size was significantly lower in the ozone-treated group at day 28 (p < 0.0001) with differences of clinical relevance (74.04% vs. 93.05% reduction of initial wound size). Histologically, the acute inflammatory reaction was enhanced in treated wounds, with significantly higher numbers of heterophils (p = 0.0016), lymphocytes (p < 0.001) and fibroblasts (p < 0.001). CONCLUSIONS: Daily topical application of ozonated sunflower oil over the course of one week improved the healing of acute, full-thickness skin wounds in chelonians. This clinical outcome was histologically correlated with an enhanced acute inflammatory reaction, as well as the production and remodelling of collagen fibres.

Effects of topical insulin on second-intention wound healing in the red-eared slider turtle (Trachemys scripta elegans) - a controlled study.

BACKGROUND: Compared with mammals, wound healing in reptiles is characterized by reduced wound contraction and longer healing times. The aim of this study is to describe the clinical and histopathological effects of topical insulin on second-intention healing of experimentally induced wounds in skin without dermal bony plates of Trachemys scripta elegans exposed to daily variations in ambient temperature and in an aquatic environment. Forty-four healthy adult females were assigned to two groups: Group 1 (n = 24) was used to assess clinical features such as wound contraction; Group 2 (n = 20) was used for histological evaluation and morphometric analysis. Topical porcine insulin (5 IU/ml diluted in glycerol) was applied daily 1 week. For each control time (2, 7, 14, 21 and 28 days post-wounding), re-epithelisation and wound remodelling were evaluated histologically and the number of main inflammatory cells (heterophils, macrophages, lymphocytes and fibroblasts) was scored. RESULTS: Mean wound contraction was higher in the insulin-treated group at each time point and differences were significant at day 28 (P < 0.0001). Histologically, these clinical findings were associated with better re-epithelisation, inflammatory response, collagen synthesis and remodelling of the wounds. Morphometrically, insulin-treated wounds had significantly higher mean counts of heterophils (day 7), macrophages (days 2, 7 and 14) and fibroblasts (days 14 and 21), whereas lymphocyte counts were significantly lower at day 21. These results demonstrate that topical insulin modifies the inflammatory response of turtle skin up-regulating inflammatory cells at early stages and promoting wound healing. CONCLUSIONS: Topical insulin is a potentially useful therapy in skin wounds of Trachemys scripta and should be evaluated in non-experimental wounds of turtles and other reptiles.

Clinical and histological findings of cutaneous wound healing in the red-eared slider turtle (Trachemys scripta elegans) housed in unheated outdoor enclosures.

BACKGROUND: Cutaneous wounds are common in chelonians. The clinical and histological features of wound healing in these species are not well described and this prevents evaluation of new therapies. OBJECTIVES: To describe clinical and histopathological features of cutaneous wound healing in the red-eared slider turtle (Trachemys scripta elegans). ANIMALS: Twenty four healthy adult females housed in outdoor facilities with free access to water and exposed to daily variations in temperature. METHODS: Full thickness 6 mm skin biopsy punch wounds were created in the rear limbs. The turtles were assigned to Group 1 (n = 12 for clinical evaluation) and Group 2 (n = 12 for microscopic study). Group 1 was photographed on Day 1 and weekly, until 28 days post wounding. Wound retraction was expressed as the percentage of perimeter reduction. For Group 2, three skin wounds were sampled at 2, 7, 14, 21, 28, 42, 60 and 135 days post wounding for histological study. The avidin-biotin-peroxidase (ABC) staining method was used to evaluate five commercial antibodies. RESULTS: Wound contraction was limited; crust persisted at least 28 days. Re-epithelialization was complete by Day 14 in many animals; active inflammation persisted until 28 days; connective tissue re-constitution and remodelling was achieved from 42 to 135 days. Antibodies AE1/AE3, Factor VIII, MAC 387, CD3 and NCL-MSA showed cross reactivity with the cell counterpart in turtle tissues. CONCLUSIONS AND CLINICAL IMPORTANCE: Second intention wound healing progressed slowly and with an indolent behaviour. Microscopically there was marked overlapping of the inflammatory and proliferative phases over a long time period.