The effectiveness of the haemodialysate Solcoseryl for second-intention wound healing in horses and ponies

Second-intention healing of limb wounds in horses is often problematic. Solcoseryl is a protein-free, standardized dialysate/ultrafiltrate (HD) derived from calf blood, which has been shown to improve healing in both animals and humans. The efficacy of HD in the healing of deep wounds in horses and ponies was investigated. Deep wounds of 20 by 35 mm were created on both metatarsi (skin, subcutis, periosteum) and on both femoral biceps muscles (skin, subcutis, muscle) of five horses and five ponies. The wounds on one side were treated with HD, four times a week during the period that the wounds were bandaged and once daily thereafter. The wounds on the other side were left untreated. In the first 4 weeks of the healing period HD stimulated healing but inhibited healing thereafter. This pattern was significant for all wound groups (P < 0.001). Because of this change in effect, the overall effect on wound healing over the entire period was not significant (P = 0.77). HD stimulated healing initially by provoking a greater initial inflammatory response, faster contraction and faster formation of granulation tissue. Subsequently, HD inhibited healing because it significantly delayed epithelialization and caused protracted inflammation. The effects of HD were most pronounced in the horses. Because this study distinguished between contraction and epithelialization, it could be shown that HD stimulated contraction but inhibited epithelialization. Therefore, HD is useful in horses for the treatment of deep wounds during the initial phase of healing by second intention, i.e. during the first weeks when wound contraction can be expected. Treatment should be ceased when epithelialization becomes predominant.     

Second-intention repair in the horse and pony and management of exuberant granulation tissue.

Second-intention repair is faster in ponies than in horses and faster in body wounds than in limb wounds. To a large extent, the differences between horses and ponies can be explained by differences in the local inflammatory response, which are a result of the functional capacity of leukocytes. In ponies, leukocytes produce more inflammatory mediators,resulting in better local defense, faster cellular debridement, and a faster transition to the repair phases, with more wound contraction. In horses,leukocytes produce fewer mediators, initiating a weak inflammatory response, which becomes chronic. This inhibits wound contraction and gives rise to the formation of exuberant granulation tissue. The anatomic environment that influences the inflammatory response and wound contraction most probably determines the differences between body and limb wounds. In body wounds, better perfusion results in faster initiation of the inflammatory phase. The weaker local resistance results in a greater degree of contraction. In limb wounds, particularly of horses, the initial inflammatory response is weak and wound contraction is restricted. Both factors give rise to chronic inflammation, which further inhibits wound contraction and promotes exuberant granulation tissue. The high incidence of exuberant granulation tissue in limb wounds of horses can thus be explained by the chronicity of the inflammatory response as well as by the common use of bandages during treatment. Chronic inflammation is often not recognized as a cause of exuberant granulation tissue. It must be prevented and treated to promote the healing process. Bandages and casts stimulate the formation of exuberant granulation tissue; however, they are advantageous in many respects and play an important role in support of the overall healing process.     

Differences in polymorphonucleocyte function and local inflammatory response between horses and ponies

REASONS FOR PERFORMING STUDY: Wound healing proceeds faster in ponies than in horses and complications during healing, such as wound infection, occur less frequently in ponies. Earlier studies suggested that this difference might be related to differences in the initial post traumatic inflammatory response. HYPOTHESIS: That polymorphonuclear leucocyte (PMN) function and profiles of humoral factors in local inflammatory processes are different in horses and ponies. METHODS: PMNs were isolated from venous blood of horses and ponies. Chemotaxis and reactive oxygen species (ROS) production was determined. Tissue cages were implanted in limbs and necks of horses and ponies and injected with carrageenan and, 3 weeks later, with LPS. In sequential samples of inflammatory exudate, the numbers of macrophages and PMNs and the production of PGE2, TNFalpha, IL-1, IL-6 and chemoattractants were determined. RESULTS: In vitro ROS production of PMNs was significantly higher in ponies than in horses, whereas in vitro PMN chemotaxis was significantly lower in ponies. In the tissue cages for both stimuli, the production of IL-1 and chemoattractants was significantly higher in ponies than in horses and remained so towards the end of the observation period in ponies. CONCLUSIONS: This study demonstrated a higher production of various inflammatory mediators by pony leucocytes. Despite the lower in vitro chemotaxis of pony PMNs, this higher in vivo production resulted in a stronger initial inflammatory response in ponies, as has been reported in studies on wound healing, through the attraction of leucocytes and triggering of the production of other cytokines. A stronger initial inflammation may promote healing by more rapid elemination of contaminants and earlier transition to repair. POTENTIAL RELEVANCE: Modulation of the initial inflammatory response might therefore be a valid option for therapeutic intervention in cases of problematic wound healing. Further, the intraspecies differences in leucocyte function may have an impact on many fields in equine medicine.     

Effect of wound location and the use of topical collagen gel on exuberant granulation tissue formation and wound healing in the horse and pony

Preformed collagen gel was topically applied to cutaneous wounds of the equine dorsal fetlock (thoracic limb) and metatarsal regions to evaluate the effect on exuberant granulation tissue production and wound healing. In 6 horses and 3 ponies (less than 140 cm high at the withers and less than 365 kg), 36 standardized cutaneous limb wounds were surgically induced (4 wounds/animal); 18 wounds were treated topically with collagen gel, and 18 wounds were not treated (controls). Collagen gel was initially applied to the wound at 0, 2, or 7 days after wound formation (groups 1, 2, and 3, respectively). Four measurements were regularly made: amount of wound contraction and the size of the granulation bed, epithelial covering, and total wound. Sequential skin and wound biopsies were evaluated histologically to assess wound healing. Using a computer, data were analyzed for differences in the 4 measurements between treated and control wounds, between fetlock wounds and metatarsal wounds, and among groups 1, 2, and 3. Analyses were performed on days 15 and 45 of wound healing and on the final day of healing. A significant difference (P greater than 0.05) in the production of exuberant granulation tissue, rate of epithelialization, or degree of wound contraction was not detected between the collagen-treated and control wounds. Total healing time and final scar size were similar. Wound healing patterns were significantly different (P less than 0.05) in the fetlock wounds and metatarsal wounds. All wounds enlarged up to day 15 with fetlock wounds enlarging significantly more than did the metatarsal wounds.(ABSTRACT TRUNCATED AT 250 WORDS)     

Differences in wound contraction between horses and ponies: the in vitro contraction capacity of fibroblasts

The contribution of wound contraction to wound closure determines the speed of second intention wound healing and it has been shown that significant differences exist with regard to both contraction and inflammatory response between horses and ponies and between various areas of the body. In this study, the contraction capacity of fibroblasts from limbs and buttocks of 4 Dutch Warmblood horses and 4 Shetland ponies was studied in vitro, in order to determine whether differences in wound contraction are due to differences in the inherent contraction capacity of the fibroblasts or to differences in tissue environmental factors, such as the inflammatory response. Fibroblasts were harvested from subcutaneous tissue, cultured and then suspended in both floating and anchored collagen gels. Contraction capacity was assessed by measuring the decrease in area of the floating gels and by measuring the microforces generated in the anchored gels using a custom-built measuring device. In the floating gels, no difference existed in the contraction capacity of fibroblasts from horses and ponies, or from limbs and buttocks. In the anchored gels, no differences existed between horse and pony fibroblasts, but the fibroblasts from the limbs started to contract significantly sooner and produced significantly higher forces than those from the buttocks. It is concluded that the in vivo differences in wound contraction between horses and ponies and between different sites of the body are not caused by differences in the inherent contraction capacity of fibroblasts. The in vitro differences between fibroblasts from limbs and buttocks are thought to be due to the lower proliferation rate and the longer culture time of the fibroblasts originating from the limbs, because mature fibroblasts can develop higher contraction forces than immature fibroblasts. This means that tissue environmental factors, such as cytokine profiles during the inflammatory response, determine the extent of contraction during wound healing. Further research should be directed towards the role of the inflammatory response in wound healing.     

Superficial Osteitis and Sequestrum Formation as a Result of Skin Avulsion in the Horse

Metatarsal injuries with exposed bone are described in seven horses. Superficial osteitis and sequestrum formation occurred in six of the seven horses. The basic pathophysiology includes local vascular stasis leading to ischemic necrosis of osteocytes and establishment of bacterial infection within dead bone. Radiographic features of superficial osteitis and sequestrum formation include reactive periosteal new bone formation, an area of radiodense necrotic bone (sequestrum) surrounded by a radiolucent zone of granulation tissue, which in turn, is surrounded by an area of increased radiodensity (involucrum). The presence of a sequestrum may usually be confirmed radiographically by 12–14 days after injury. Surgical management included wound debridement, wound irrigation, bandaging or casting, sequestrectomy and skin grafting. Five of the seven patients required sequestrectomy, in one the sequestrum disappeared without removal. Mixed bacterial populations were commonly found in the wounds. No specific etiologic agent was associated with sequestrum formation.     

Comparison of equine amnion and a nonadherent wound dressing material for bandaging pinch-grafted wounds in ponies

OBJECTIVE: To evaluate healing of pinch-grafted wounds on the distal aspect of the limbs of ponies bandaged with equine amnion or a standard nonadherent wound dressing material. ANIMALS: 6 ponies. PROCEDURE: A 2.5x2.5-cm full-thickness section of skin was removed from the dorsal aspect of each limb at the midpoint of the metacarpus or metatarsus. Six days later, wounds were grafted with partial-thickness pinch grafts. Half the wounds were bandaged with amnion, and the other half were bandaged with a nonadherent dressing. Bandages were changed every 3 days until wound healing was complete. At each bandage change, numbers of grafts lost were recorded, and wounds were measured. RESULTS: Percentage of grafts lost from wounds bandaged with amnion was not significantly different from percentage lost from wounds bandaged with the nonadherent dressing. Median healing time for wounds bandaged with amnion (30 days) was significantly less than median healing time for wounds bandaged with the nonadherent dressing (39 days). All wounds were healed by day 45. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that amnion can be used for bandaging pinch-grafted wounds on the distal aspect of the limbs of ponies.     

Delayed Wound Closure A Review and Report of Use of the Technique on Three Equine Limb Wounds

Limb wounds of three horses were managed by delayed wound closure. In each, the wound was such that it was unlikely that healing would occur following primary closure and that second intention healing would have been unsatisfactory. Wounds were cleaned and debrided at the time of admission and maintained under sterile pressure bandages until free of infection and devitalized tissue. Suture closure of the wound at that time resulted in satisfactory healing in each case.     

Wounds of the pastern and foot region managed with phalangeal casts: 50 cases in 49 horses (1995–2006)

Objective   Describe the use of a phalangeal cast as treatment for wounds in the pastern and foot region of horses. Secondly, to evaluate the healing and soundness of horses treated with phalangeal casts.
Design   Retrospective study of 49 horses.
Procedures   Medical records of 49 horses that were treated with a phalangeal cast for 50 cases of wounds in the pastern and foot region at equine referral hospitals from 1995 to 2006 were reviewed and follow-up information was obtained.
Results   Treatment consisted of wound debridement, lavage, wound closure (28 wounds), cast application and antibiotics (84%). At follow-up, the majority of horses were sound (42 of 47 wounds, 89.4%), three horses were still lame and one horse was euthanased because of persistent lameness. Three horses were lost to follow-up. There was no statistical difference between the outcomes of horses treated acutely (<24 h) or after a 24-h delay. Similarly, the involvement of synovial structures in the wound did not significantly influence outcome.
Conclusions   In this study, wounds involving the pastern and foot that were treated with a phalangeal cast carried a good prognosis for soundness (89.4%) and cosmetic healing (89.5%). The phalangeal casts were well-tolerated and effective.     

The effects of topical oxygen therapy on equine distal limb dermal wound healing

Topical oxygen therapy (TOT) has been used in human medicine to promote healing in chronic wounds. To test the efficacy and safety of TOT in horses, an experimental wound model was created by making 1 standardized dermal wound on each limb of 4 healthy horses (n = 16). Each wound was fitted with an oxygen delivery cannula and covered with a bandage. One limb of each front and hind pair was randomly assigned to the treatment group (fitted with an oxygen concentrator device), with the contralateral limb assigned to the control group (no device). Wound area, epithelial area, and contraction were measured every 3 to 4 d. Biopsy samples and culture swabs were taken on days 16 and 32 to evaluate angiogenesis, fibroplasia, epithelial hyperplasia, inflammation and bacterial growth. Mean healing time in treated wounds (45 d, range: 38 to 52 d) was not significantly different from that in the paired control wounds (50 d, range: 38 to 62 d). Topical oxygen therapy had little effect on dermal wound healing in this experimental wound model in healthy horses.     

Application of Exogenous Esterified Hyaluronan to Equine Distal Limb Wounds

The objective of this study was to evaluate the efficacy of topical application of a hyaluronan (HA) derivative in wound healing with respect to the rate of epithelialization, fibroplasia, angiogenesis and contraction, magnitude of the local inflammatory response, local expression of transforming growth factor-β 1 and 3 (TGF-β 1 and 3), tumor necrosis factor-α (TNF-α), and collagen type III deposition. In six healthy adult horses, six full-thickness skin wounds were created on the dorsal aspect of both metacarpi using a sterile template. Sites were sampled at 0, 1, 2, 5, 14, 21, and 35 days following wounding. Wounds on one limb were dressed with commercially available esterified HA fleece under a nonadherent dressing. The opposite limb was covered with the nonadherent dressing alone (control). Images of the most proximal wounds were used to determine the area of total healing and the relative contributions of epithelialization and contraction to healing. At each sample time, a control and treatment biopsy were taken for histological evaluation and special stains. All samples were evaluated for degree of inflammation, fibroplasia and angiogenesis; in situ hybridization for type III collagen, TGFβ1 and 3, and immunohistochemistry for TNF-α. Mean percentages of total wound healing, epithelialization, and wound contraction were not significantly different between control and treatment groups. In treated horses, initial wound expansion was significantly decreased during the first 2 weeks. Mononuclear cell numbers, counted in the granulation tissue, increased in both control and treated limbs over the entire course of the study. However at day 35 the macrophage numbers counted in the treated horses were significantly increased compared with the control limbs (P < .05). Although not statistically significant, relative staining for type III collagen in the treated wounds was less than that of control wounds. Results of the present study do not support a benefit of an exogenous HA-derivative in the healing of distal limb wounds in horses. The shortcomings of the study design are discussed.     

Cystometrography and urethral pressure profiles in healthy horse and pony mares

Cystometrography and urethral pressure evaluations were performed in 7 horse mares and 5 pony mares before and after sedation with xylazine. Before sedation, mean (+/- SD) maximal bladder contraction pressure was 91.4 +/- 16.5 cm of H2O in horses and was 86.0 +/- 14.4 cm of H2O in ponies, and maximal urethral closure pressure was 49.1 +/- 19.4 cm of H2O in horses and 37.7 +/- 14.4 cm of H2O in ponies. A significant difference was not found between values of nonsedated vs sedated animals. Only values for threshold volume were significantly different (P less than 0.05) between nonsedated horses (1,982 +/- 1,241 ml) and ponies (825 +/- 412 ml).     

Growth characteristics of fibroblasts isolated from the trunk and distal aspect of the limb of horses and ponies

OBJECTIVE: To determine if there is a difference in in vitro growth of fibroblasts isolated from the trunk and distal aspect of the limb of horses and ponies. To determine the effects of a corticosteroid and monokine on in vitro growth of fibroblasts isolated from the trunk and distal aspect of the limb of horses and ponies. STUDY DESIGN: Growth of fibroblasts from tissues harvested from the trunk and limb were compared from horse and pony samples grown in control media and control media with triamcinolone or monokine added. ANIMALS OR SAMPLE POPULATION: Dermal and subcutaneous tissue from 22 horses and 17 ponies of various ages and breeds. METHODS: Fibroblast growth was assessed by tritiated thymidine uptake using standard cell culture techniques. The effect of a monokine or triamcinolone plus control media were compared with control media for fibroblast growth. RESULTS: Fibroblast growth from tissues isolated from the horse limb was significantly less than growth from the horse trunk and the limb and trunk of ponies. Monokine was more effective than triamcinolone in suppressing fibroblast growth from tissues isolated from the trunk and limb in both horses and ponies. CONCLUSIONS: There are growth differences in fibroblasts isolated from the limb of horses compared with those isolated from the trunk and from the limb and trunk of ponies. CLINICAL RELEVANCE: The difference in fibroblast growth from tissues isolated from the trunk and limb of horses and ponies may provide evidence for the difference reported in the healing characteristics of limb wounds in horses and ponies. Influencing fibroblast growth may provide a key to controlling the development of exuberant granulation tissue in horses and ponies.     

Negative-Pressure Wound Therapy as Management of a Chronic Distal Limb Wound in the Horse

Wound healing in distal limb wounds with tissue loss in horses may be an expensive and long process due to prolonged inflammatory phase. Negative pressure wound therapy (NPWT) (also known as vacuum-assisted closure) is well established method of wound therapy in human plastic and reconstructive surgery for many years. It consists of a leak free bandage and a pump that applies subatmospheric pressure to the wound area. This report describes the successful use of the NPWT intended for the suppression of exuberant granulation tissue development of the wound on distal limb in a horse.     

The modified Meek technique as a novel method for skin grafting in horses: evaluation of acceptance, wound contraction and closure in chronic wounds

REASONS FOR PERFORMING STUDY: The acceptance of skin grafts in horses is unpredictable and the final cosmetic result can be disappointing. Besides movement and infection, graft failure is often caused by chronic inflammation, inherently present during second intention healing of limb wounds in horses. In human burns affected by infection and inflammation, the acceptance of the island skin grafts of the modified Meek technique appeared to be better than meshed sheet skin grafts. HYPOTHESIS: The percentage take of Meek micrografts is higher than of other techniques; and rates of both wound contraction and epithelialisation are increased. METHODS: Large traumatic limb wounds of 13 horses healing by second intention were grafted using the modified Meek technique. Photographs of the wounds were taken at set intervals. Wound areas, and areas of acceptance and rejection were determined using a digital image post processor (Scion Image). The percentages of take, wound contraction and epithelialisation were calculated. RESULTS: The initial mean wound area was 7500 mm2. Graft acceptance was mean +/- s.d. 93.7 +/- 5.9%. Wound closure was due to contraction (55.2 +/- 11.1%) and epithelialisation (44.8 +/- 11.1%) and resulted in a 96.7 +/- 3.6% reduction of the initial wound area 29.1 +/- 6 days after grafting. All wounds showed functional and cosmetic healing. CONCLUSIONS: The method for skin grafting in horses achieved higher percentages of take than reported previously and consistent cosmetic and functional results. The grafts increased not only the rate of epithelialisation but also had a strong positive effect on wound contraction, resulting in rapid closure and smaller scars. POTENTIAL RELEVANCE: The modified Meek technique proved to be a novel technique for skin grafting equine wounds in clinical practice, which can be performed easily. The molecular background of the increase of wound contraction by the grafts may provide a clue in the search for medicinal stimulation of wound contraction during second intention healing.     

Effects of topical application of antimicrobials and bandaging on healing and granulation tissue formation in wounds of the distal aspect of the limbs in horses

OBJECTIVE: To determine whether povidone iodine ointment or 2 forms of silver sulfadiazine applied topically to wounds of the distal aspect of the limbs in horses affect the rate of second intention healing and to evaluate the additional influence of bandaging with these antimicrobials on granulation tissue formation. ANIMALS: 6 healthy adult horses. PROCEDURE: Six standardized 2.5-cm2 skin wounds/horse were distributed between the dorsomedial surfaces of the metacarpi and metatarsi. One of the following 6 treatments was applied to each wound: 1% silver sulfadiazine cream with bandage, 1% silver sulfadiazine slow-release matrix with bandage, 1% silver sulfadiazine slow-release matrix without bandage, povidone-iodine ointment with bandage, untreated control with bandage, and untreated control without bandage. Wound area, granulation tissue area, and perimeter were measured by use of planimetry software applied to digital images. Exuberant granulation tissue was excised when present. Days until healing, rate of healing parameter, rate of contraction, and epithelialization were compared among wound treatment groups. RESULTS: Healing parameters and mean days to healing did not differ significantly among any of the wound treatment groups. Percentage wound contraction and rate of epithelialization were similar among wound treatments. All bandaged wounds produced exuberant granulation tissue, which was surgically excised; none of the unbandaged wounds produced exuberant granulation tissue. CONCLUSIONS AND CLINICAL RELEVANCE: When exuberant granulation tissue is removed, rates of epithelialization and wound contraction were not different among wound treatment groups, whether bandaged or unbandaged. Topical application of 1% silver sulfadiazine slow-release matrix on unbandaged wounds induced the same result as medications applied beneath bandages, but without exuberant granulation tissue formation.     

Use of negative pressure wound therapy in three horses with open,infected olecranon bursitis

This article describes the clinical use of negative pressure wound therapy (NPWT) in three horses with an open and infected subcutaneous olecranon bursa, in which prior surgical excision followed by primary closure was unsuccessful. The wounds filled in rapidly with granulation tissue and contracted during therapy. All wounds subsequently went on to heal without complications. The use of NPWT was well tolerated in all three horses and facilitated healing in a difficult area.     

Effects of Topical Application of Sunflower-Seed Oil on Experimentally Induced Wounds in Horses

The objectives were to evaluate clinical and histopathological aspects of topical application of sunflower-seed oil (Helianthus annuus) on the healing process of experimentally induced wounds in lumbar and metacarpal areas of healthy horses. Six adult horses were used. Eight wounds were surgically produced on each horse: two 6.25-cm² wounds cranial to the sacrum on each side of the lumbar region and two 2.89-cm² wounds close to the proximal epiphysis of the metacarpus on the lateral aspect of each forelimb. Left side was used for macroscopic observations and right side for histopathological analysis. The experimental lesions were treated daily with sunflower-seed oil, whereas saline solution was used in control lesions. Macroscopic and histopathological analyses were performed on tissue harvested at 3, 7, 14, and 21 days. Complete healing time for all wounds was recorded. For lumbar region’s wounds, a contraction of 90.78% was recorded for those treated with oil and of 79.27% for control wounds after 21 days of treatment. For metacarpal region’s wounds, a contraction of 47.63% was recorded for wounds treated with oil and of 30.21% for control wounds. Wounds in the sunflower-seed oil treatment group had an elevation of polymorphonuclear cells, a newly formed vascular bed during the inflammatory phase, and a better alignment of collagen fibers during the remodeling phase. In conclusion, topical application of sunflower-seed oil was beneficial in the healing process of experimentally induced skin wounds in horses, with best results for treatment of lumbar wounds, making it a therapeutic option in equine wound healing.     

Partial Arytenoidectomy in the Horse with and without Mucosal Closure

Left-sided partial arytenoidectomy was performed in eight horses to evaluate healing. Four horses underwent conventional partial arytenoidectomy with suture apposition of the mucosa. In four horses, most of the arytenoid cartilage, including overlying mucosa, vocal fold, and laryngeal saccule, were excised en bloc without mucosal closure. The horses were monitored clinically by endoscopic examination. One horse from each group was euthanatized at weeks 2, 4, 8, and 16. Complete necropsies with gross and histologic examination of the arytenoidectomy sites were performed. Postoperative complications such as coughing, dysphagia, and aspiration pneumonia were not encountered and problems with wound healing were minimal in both groups. The defect created by partial arytenoidectomy without mucosal closure initially filled to the level of the luminal surface with granulation tissue, with a gradual transition to mature fibrous connective tissue. Grossly, the defect appeared to be healed and was completely epithelialized by week 16 without apparent narrowing of the laryngeal lumen. Partial arytenoidectomy sites with mucosal closure healed in a similar pattern by week 8. Partial dehiscence of the dorsal portion of the sutured mucosa occurred in three horses.